Hydrogels are remarkable materials known for their ability to absorb and retain large amounts of water. The type of hydrogels used for atmospheric water harvesting, like the one developed by MIT, are specifically engineered to not only absorb water vapor from the air but also to efficiently release it for collection as liquid water.
Here’s a breakdown of the kind of hydrogels that can absorb and “make” water in this context:
1. Hydrophilic Polymer Networks:
At their core, hydrogels are three-dimensional networks of hydrophilic (water-attracting) polymers. These polymers are cross-linked, meaning they are chemically or physically bonded together to form a stable, insoluble gel structure.
The hydrophilic groups (like hydroxyl, carboxyl, or amine groups) within the polymer chains have a strong affinity for water molecules, allowing the gel to swell significantly as it absorbs moisture.
To enhance their water absorption capabilities, especially in dry air, hydrogels used for atmospheric water harvesting are often infused with hygroscopic salts, such as lithium chloride (LiCl).
Salts like LiCl are powerful desiccants, meaning they naturally attract and hold water molecules from the surrounding environment, even at low humidity levels.
MIT’s research, in particular, focused on significantly increasing the amount of lithium chloride infused into the hydrogel (up to 24 grams of salt per gram of polymer) to achieve “record-breaking” vapor absorption.
3. Tuned Microstructure and Composition for Absorption and Release:
Swelling and Deswelling: The hydrogels are designed to undergo reversible swelling and deswelling. They absorb water vapor from the air, expanding like a sponge. When heated (e.g., by sunlight), they release the absorbed water as vapor.
Preventing Salt Leakage: A key innovation in the MIT hydrogel is the incorporation of glycerol. Glycerol is a liquid compound that helps stabilize the lithium salt within the hydrogel, preventing it from crystallizing and leaking out with the collected water. This ensures the collected water is safe for drinking, as salt levels remain below standard thresholds.
Designed Porosity/Structure: While some hydrogels have micro or nano-pores that can lead to salt leakage, the MIT design specifically engineered the hydrogel’s microstructure to lack nanoscale pores, further limiting salt escape. Additionally, they molded the hydrogel into a “bubble wrap”-like pattern of small domes to increase surface area, optimizing water vapor absorption and release.
Thermo-responsiveness: These “smart” hydrogels are often thermo-responsive, meaning their ability to absorb and release water is influenced by temperature. The MIT hydrogel, for instance, releases water efficiently when heated by the sun. Some hydrogels, like polyethylene glycol (PEG), have even shown an increase in water absorption as temperatures climb, due to a phase transformation in their microstructure.
In essence, the hydrogels capable of absorbing and “making” water are sophisticated polymeric materials, often enhanced with specific salts and designed with optimized structures to efficiently capture atmospheric moisture and then release it as clean liquid water through a controlled process, often driven passively by solar heat.
Methylene Blue, also known by its chemical name methylthioninium chloride and commonly referred to as Swiss Blue, is a versatile compound with a rich history spanning over a century. Initially synthesized in 1876 by the German chemist Heinrich Caro, its primary application was as an aniline-based dye within the textile industry. The subsequent discovery of its unique properties led to its adoption in various scientific and medical fields, marking it as the “first fully synthetic drug used in medicine”. This report aims to provide a comprehensive overview of Methylene Blue, encompassing its chemical identity, fundamental properties, historical and current medical uses, ongoing scientific research into potential future applications, its diverse non-medical uses, potential health benefits that are still under investigation, the known risks and side effects associated with its use, and its legal and regulatory status within the United States.
2. Chemical Identity and Fundamental Properties
2.1 Chemical Formula and Nomenclature
The chemical formula for Methylene Blue is It is important to note that Methylene Blue can also exist in a hydrated form, which contains three molecules of water per unit of the compound. The International Union of Pure and Applied Chemistry (IUPAC) name for Methylene Blue is [7-(dimethylamino)phenothiazin-3-ylidene]-dimethylazanium;chloride. Beyond these formal designations, Methylene Blue is known by a variety of other common names and synonyms, including Methylthioninium chloride, Swiss Blue, Basic Blue 9, CI 52015, Urelene blue, Provayblue, Proveblue, and Methylenium ceruleum. Chemically, Methylene Blue is classified as a formal derivative of phenothiazine and belongs to the thiazine dye family. The existence of multiple names and the distinction between anhydrous and hydrated forms underscore the necessity for precision when referring to or utilizing this compound in both research and clinical settings. Subtle differences in these forms, such as solubility and stability, can influence its behavior and efficacy.
2.2 Molecular Weight and Structure
The molar mass of anhydrous Methylene Blue is approximately 319.85 g/mol, while the trihydrate form has a molar mass of 373.9 g/mol. The molecular structure of Methylene Blue is characterized by three interconnected cyclic structures. A central phenothiazine ring system is linked to sulfur and nitrogen atoms, featuring dimethylamino groups at positions 3 and 7. The molecule carries a positive charge on a nitrogen atom, which is balanced by a chloride counterion. This structural arrangement classifies Methylene Blue as a cationic heterocyclic compound. A key feature of Methylene Blue is its redox activity, which allows it to exist in two primary forms: an oxidized state, which is blue in color, and a reduced state, known as leuco-methylene blue, which is colorless. This ability to readily accept and donate electrons is fundamental to many of its biological activities, including its established use in treating methemoglobinemia and its potential role in influencing mitochondrial function.
2.3 Physical and Chemical Properties
At room temperature, Methylene Blue presents as a dark green crystalline powder, often exhibiting a bronze-like luster. When dissolved in polar solvents such as water or alcohol, it yields a characteristic deep blue solution. Its solubility varies across different solvents: it is readily soluble in water, glycerol, chloroform, glacial acetic acid, and ethanol; slightly soluble in pyridine; and practically insoluble in ethyl ether, oleic acid, and xylene. The specific solubility in water is reported to be around 4.36 g per 100 mL at 25°C. Notably, the solubility in solvents like DMSO and ethanol can be enhanced at elevated temperatures. The melting point of Methylene Blue is in the range of 100 to 110 °C, at which point it also begins to decompose , although some sources indicate a decomposition temperature closer to 180°C. When dissolved in water, Methylene Blue exhibits slightly acidic properties , with a 1% aqueous solution having a pH of approximately 6. The compound absorbs light maximally in the region of 664 to 670 nm. While generally stable under normal conditions, Methylene Blue is sensitive to light, which can lead to its degradation. The solubility profile of Methylene Blue is a critical factor influencing its administration and distribution within biological systems. Its light sensitivity has implications for its use in photodynamic therapy and necessitates careful storage to maintain its potency [Chain of thought: How it dissolves affects how it can be formulated (e.g., for injection vs. oral). Light degradation could impact the potency of stored solutions.].
Table 1: Chemical and Physical Properties of Methylene Blue
Methylthioninium chloride, Swiss Blue, Basic Blue 9, CI 52015, etc.
Molecular Weight (Anhydrous)
319.85 g/mol
Molecular Weight (Trihydrate)
373.9 g/mol
Appearance
Dark green crystalline powder with a bronze-like luster
Solubility in Water
4.36 g/100 mL at 25°C
Solubility in Other Solvents
Soluble in glycerol, chloroform, glacial acetic acid, ethanol; slightly soluble in pyridine; insoluble in ethyl ether, oleic acid, xylene
Melting Point
100 to 110 °C (decomposes)
Maximum Absorption Wavelength
664-670 nm
3. A Historical Perspective on Methylene Blue in Medicine
3.1 Early Discoveries and Applications
The journey of Methylene Blue from a textile dye to a significant medical agent began with its recognition as a valuable biological stain. In 1880, Robert Koch, a pioneer in microbiology, established its utility as a stain in medical applications, a finding corroborated and expanded upon by Paul Ehrlich. Ehrlich, in the 1890s, made a groundbreaking observation: Methylene Blue exhibited effectiveness against malaria parasites in human blood, marking it as the first fully synthetic drug to be successfully used in treating human illnesses. This discovery aligned with Ehrlich’s “magic bullet” theory, which posited that specific chemicals could selectively target and harm pathogens without damaging surrounding tissues, a revolutionary concept that laid the foundation for modern chemotherapy. During World War I, Methylene Blue also found application as an antiseptic for treating wounds, demonstrating its antimicrobial properties. It was, in fact, the first synthetic antiseptic to be used therapeutically. Historically, it was also employed in the treatment of gonorrhea and fever. These early applications highlight the initial promise and versatility of Methylene Blue in addressing various medical challenges.
3.2 Treatment of Methemoglobinemia
A significant milestone in the medical history of Methylene Blue occurred in 1933 when it was discovered to be an effective antidote for aniline-induced methemoglobinemia by Williams and Challis. Methemoglobinemia is a condition characterized by an elevated level of methemoglobin in the blood, a form of hemoglobin that cannot effectively carry oxygen to the body’s tissues. Methylene Blue works by chemically reducing the ferric iron (Fesup3+/sup) present in methemoglobin back to the ferrous iron (Fesup2+/sup) state in hemoglobin, thereby restoring the blood’s oxygen-carrying capacity. This mechanism has established Methylene Blue as a crucial treatment for acquired methemoglobinemia, which can be caused by exposure to certain pharmaceuticals, toxins, or even broad beans in susceptible individuals. The effectiveness of Methylene Blue in this context is a testament to its direct and specific biochemical action on hemoglobin.
3.3 Other Historical Uses
Beyond its roles in malaria treatment and methemoglobinemia, Methylene Blue has been explored for various other medical applications throughout history. It was once considered a weak antimalarial agent, but its use diminished with the advent of more potent drugs like chloroquine. However, the increasing prevalence of drug-resistant malaria has led to a renewed interest in Methylene Blue as a potential component of antimalarial treatment regimens. Similarly, Methylene Blue was historically recommended as an intestinal and urinary antiseptic, although this use is no longer prevalent. Nevertheless, some sources still mention its application in treating urinary tract infections. In 1933, Matilda Brooks discovered its potential as an antidote for both cyanide and carbon monoxide poisoning , although it is no longer the primary treatment for cyanide poisoning. Notably, Methylene Blue was also one of the first drugs used in the late 19th century for the treatment of patients with psychosis and played a role in the serendipitous development of phenothiazine antipsychotic drugs in the mid-20th century. The varied trajectory of Methylene Blue’s medical applications reflects the continuous advancements in pharmacological science and the development of more targeted therapies. The resurgence of interest in its antimalarial properties highlights the ongoing challenges posed by drug resistance.
4. Current FDA-Approved Medical Applications of Methylene Blue
4.1 Treatment of Acquired Methemoglobinemia
The primary FDA-approved medical application of Methylene Blue is the treatment of acquired methemoglobinemia in both pediatric and adult patients. This approval underscores the established efficacy and safety of Methylene Blue for this specific condition. Several intravenous formulations are available, including ProvayBlue, which was the first methylene blue injection to receive FDA approval. Generic versions of methylene blue injection have also been approved by the FDA, enhancing the accessibility of this essential medication. The typical intravenous dosage for treating methemoglobinemia is 1 mg/kg of a 1% solution, administered slowly over a period of 5 to 30 minutes. If methemoglobin levels remain elevated or clinical signs persist, a repeat dose may be administered one hour after the initial dose. The FDA approval of Methylene Blue for methemoglobinemia signifies a robust body of evidence supporting its benefit in this critical medical situation.
4.2 Diagnostic Aid
Methylene Blue is also FDA-approved for various diagnostic purposes, leveraging its staining properties to enhance visualization during medical procedures. One significant application is in sentinel lymph node mapping, a crucial technique used during breast surgery and other cancer surgeries to identify the lymph nodes most likely to contain cancerous cells. In this procedure, a typical dose of 2 mL or 5 mL of a 1% methylene blue solution is injected into the tissue near the tumor, allowing surgeons to visually trace the lymphatic drainage. Furthermore, Methylene Blue is employed as a dye in chromoendoscopy, where it is sprayed onto the mucosa of the gastrointestinal tract to aid in the identification of dysplasia, or pre-cancerous lesions. It is also utilized in endoscopic polypectomy as an adjunct to saline or epinephrine injection. Another diagnostic application involves the intravenous administration of Methylene Blue to assist in the identification of parathyroid glands during surgery. The dye stains the glands, making them easier for surgeons to locate. A typical dose for this purpose is 5 mg/mL given approximately one hour before the surgical procedure. Additionally, because intravenously injected Methylene Blue is readily excreted in the urine, it can be used to test for leaks or fistulas within the urinary tract. These diagnostic uses highlight the value of Methylene Blue’s staining properties in improving the precision and effectiveness of various medical and surgical procedures.
4.3 Combination Drug for Urinary Tract Issues
Methylene Blue is also a component of several FDA-approved combination drugs, such as Hyophen, Methylphen, Urophen, and Urised. These medications are indicated for the symptomatic treatment of pain, burning, urgency, and frequency associated with cystitis, urethritis, and other urinary tract disorders. In these formulations, Methylene Blue is combined with other active ingredients like hyoscyamine, hexamethylenetetramine, phenyl salicylate, and benzoic acid. While Methylene Blue has a history of use as a mild urinary antiseptic, its current approved application in this context is within these multi-component drugs, suggesting a synergistic or complementary role in alleviating urinary tract symptoms.
Table 2: Current FDA-Approved Medical Applications of Methylene Blue
Indication
Route of Administration
Typical Dosage
Specific FDA Approval Details
Acquired Methemoglobinemia
Intravenous
1 mg/kg of a 1% solution over 5-30 minutes, repeat dose if needed
ProvayBlue (first approved brand), generic versions available
Sentinel Lymph Node Mapping
Intraparenchymal
2 mL or 5 mL of a 1% solution injected near the tumor
Used in breast surgery and other cancer surgeries
Visualization in Endoscopic Procedures
Topical (spray/injection)
Varies depending on the procedure
Chromoendoscopy for dysplasia detection, endoscopic polypectomy
Parathyroid Gland Identification
Intravenous
5 mg/mL one hour before the procedure
Aids in locating parathyroid glands during surgery
Urinary Tract Leak Detection
Intravenous
Dosage not specified
Excreted in urine to detect leaks or fistulas
Symptomatic Treatment of Urinary Tract Pain
Oral (in combination drugs)
Dosage determined by the specific combination product
Component of drugs like Hyophen, Methylphen, Urophen, Urised, which also contain hyoscyamine, hexamethylenetetramine, phenyl salicylate, and benzoic acid
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5. Ongoing Scientific Studies and Potential Future Medical Uses of Methylene Blue
5.1 Neurodegenerative Disorders
Methylene Blue has garnered significant attention for its potential in treating various neurodegenerative disorders. Extensive research is underway to explore its effects on Alzheimer’s disease, with studies investigating its ability to inhibit the aggregation of tau proteins and reduce the formation of amyloid-beta plaques, both hallmarks of the disease. Some clinical trials, particularly those involving modified forms of Methylene Blue like LMTX (leuco-methylthioninium bis(hydromethanesulfonate)), have shown promising results in specific subgroups of patients. Lower doses of Methylene Blue have also been associated with enhanced cognitive function in some studies. However, it is important to note that several large-scale phase 3 clinical trials have not met their primary endpoints, indicating the need for further investigation to optimize dosing, formulations, and identify the patient populations that might benefit most. Preclinical studies also suggest that Methylene Blue may have neuroprotective effects in Parkinson’s disease by reducing oxidative stress and protecting dopaminergic neurons. Furthermore, its potential is being explored in other neuropsychiatric disorders, including bipolar disorder, claustrophobia, ifosfamide encephalopathy, and schizophrenia , as well as autism, depression, neurodegenerative diseases, and traumatic brain injury. A completed clinical trial indicates that intraoperative use of Methylene Blue may reduce postoperative delirium and cognitive dysfunction in elderly patients undergoing major noncardiac surgery. The ability of Methylene Blue to cross the blood-brain barrier and its potential to enhance mitochondrial function and act as an antioxidant are key reasons for its investigation in these neurological conditions [Chain of thought: While preclinical data is promising, translating these findings to consistent clinical benefits in complex neurodegenerative diseases is challenging and requires rigorous investigation.].
5.2 Cancer Therapy
The role of Methylene Blue in cancer therapy is also an active area of research. Its potential in photodynamic therapy (PDT) is being explored as a means to selectively kill cancer cells. Preclinical studies have shown its effectiveness against various cancer types, including colorectal tumors, carcinoma, and melanoma , although results have been less promising in breast cancer and HeLa cell models. Clinical trials are investigating its efficacy in treating pain associated with oral mucositis in cancer patients. Researchers are also examining its potential to enhance the effectiveness of radiation therapy, particularly in making hypoxic tumor cells more susceptible to radiation. In animal models, Methylene Blue has shown promise as a metabolic therapy in restraining ovarian tumor growth. Additionally, its use in facilitating the evaluation of lymph nodes in colon cancer specimens is being studied. The photosensitizing properties of Methylene Blue, activated by light to produce cytotoxic singlet oxygen, and its potential to interfere with cancer cell metabolism are the basis for these investigations.
5.3 Infectious Diseases
There is a renewed interest in Methylene Blue as an antimalarial agent, particularly in the face of increasing resistance to existing drugs. Studies are exploring its effectiveness against drug-resistant strains of malaria and its ability to prevent transmission by targeting the gametocyte stage of the parasite. Methylene Blue is also being investigated for its broad-spectrum antiviral activity, with potential applications against respiratory viral infections such as influenza and SARS-CoV-2. Experimental studies have demonstrated its inhibitory effects on viral replication. Its antimicrobial properties are also being researched in the context of treating bacterial infections, including urinary tract infections, and as a general disinfectant. Notably, laboratory studies suggest its effectiveness against persister biofilms, which are relevant to chronic infections like Lyme disease and Bartonella. Furthermore, Methylene Blue is used in some settings for decontaminating blood plasma products due to its antiviral and antibacterial properties. The diverse antimicrobial and antiviral activities of Methylene Blue, combined with its relatively low toxicity and cost, make it a promising candidate for addressing infectious diseases, especially in resource-limited settings and against emerging pathogens.
5.4 Septic Shock and Vasoplegic Syndrome
Methylene Blue is under investigation for its potential to treat refractory hypotension in septic shock. Its mechanism of action involves inhibiting nitric oxide synthase and guanylate cyclase, which can help restore vascular tone. Clinical trials are currently ongoing to further evaluate its safety and efficacy in this context. Additionally, Methylene Blue is used off-label to increase blood pressure in individuals experiencing vasoplegic syndrome, a condition often occurring after cardiac surgery where blood pressure drops dangerously low and does not respond to standard treatments like epinephrine. Some studies suggest that early administration of Methylene Blue in patients with vasoplegic syndrome may improve survival rates. The ability of Methylene Blue to modulate nitric oxide pathways, which play a critical role in vasodilation, makes it a potential therapeutic option for managing these severe conditions.
Table 3: Potential Future Medical Uses of Methylene Blue Under Investigation
Medical Condition
Proposed Mechanism of Action
Current Status of Research
Key Findings/Outcomes (if available)
Alzheimer’s Disease
Inhibits tau aggregation, reduces amyloid-beta plaques, enhances mitochondrial function
Preclinical and Clinical Trials (Phase 2/3)
Mixed results in clinical trials, some promise in subgroups with modified forms and lower doses
Demonstrates antiviral activity in laboratory settings
Bacterial Infections (e.g., Lyme, Bartonella)
Antimicrobial, effective against persister biofilms
Preclinical and Anecdotal Reports
Shows promise in laboratory studies and some clinical observations
Septic Shock
Inhibits nitric oxide synthase and guanylate cyclase, restores vascular tone
Clinical Trials (Phase 2/3)
Some studies show improved hemodynamic parameters and reduced vasopressor requirements
Vasoplegic Syndrome
Inhibits nitric oxide synthase and guanylate cyclase, restores vascular tone
Off-label use, Retrospective and Prospective Studies
Early administration may improve survival after cardiac surgery
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6. Explore Non-Medical Applications of Methylene Blue
6.1 Use in Aquariums
Methylene Blue finds application in the maintenance of aquariums, primarily as a disinfectant. It is commonly used to treat fungal infections that can affect fish and their eggs, as well as parasitic diseases such as ich (Ichthyophthirius multifiliis). Additionally, it can be effective against certain bacterial infections in fish. Beyond treating existing conditions, Methylene Blue can also aid in mitigating the toxicity caused by elevated levels of ammonia and nitrites in aquarium water, which can be harmful to aquatic life. Its antimicrobial properties, therefore, extend beyond medical contexts to play a role in maintaining the health and well-being of fish in aquariums.
6.2 Dye in Textiles and Microscopy
Historically, Methylene Blue was first synthesized for use as a dye in the textile industry, particularly for coloring cotton and wool fabrics. Its strong and lasting blue hue made it a valuable component in textile manufacturing. In the realm of science, Methylene Blue is a widely employed biological stain in microscopy. It is used to enhance the visibility of cells, tissues, and microorganisms under a microscope, often staining negatively charged cell components such as nucleic acids. It is a component of important stains used in hematology and microbiology, including Wright’s stain and Jenner’s stain. The fundamental staining properties of Methylene Blue were crucial to its early scientific and industrial applications, highlighting its inherent affinity for biological materials.
6.3 Redox Indicator in Chemistry
Methylene Blue is a well-known redox indicator in analytical chemistry. Solutions containing Methylene Blue will appear blue in the presence of an oxidizing environment. However, if exposed to a reducing agent, the solution will undergo a color change and become colorless as the Methylene Blue is reduced to its leuco form. This property is famously demonstrated in the “blue bottle” experiment, a classic demonstration of chemical kinetics. Methylene Blue is also utilized in sulfide analysis, where its reaction with hydrogen sulfide can be quantified. Additionally, it can function as an indicator for pH changes in certain applications. The reversible color change of Methylene Blue based on the redox state of its environment makes it a valuable tool for visualizing and studying chemical reactions involving electron transfer.
6.4 Photosensitizer
Methylene Blue acts as a photosensitizer, meaning it can absorb light and transfer energy to oxygen, converting it into singlet oxygen, a highly reactive form. This property is harnessed in photodynamic therapy (PDT), where Methylene Blue, in conjunction with light exposure, is used to destroy target cells, such as cancer cells or microorganisms. The same principle is applied in the disinfection of blood plasma, where light-activated Methylene Blue can effectively kill certain viruses and bacteria, enhancing the safety of blood transfusions. This interaction with light to generate reactive oxygen species is a key aspect of its potential in targeted therapeutic applications.
6.5 Other Uses
Beyond these primary non-medical applications, Methylene Blue has a variety of other uses. In the food industry, it is employed to test the freshness of milk and dairy products, where its reduction to a colorless form indicates low oxygen levels associated with spoilage. Its use in the textile industry for dyeing natural fibers like cotton, wool, and silk continues. In educational settings, Methylene Blue is a valuable tool for demonstrating redox reactions and chemical equilibrium, such as in the Blue Bottle Experiment, and for staining cells to facilitate microscopic observation by students. It also has applications in environmental science, where it can be used to demonstrate dissolved oxygen levels in water, illustrating concepts of water quality. In orthopedic surgery, Methylene Blue is sometimes added to bone cement to provide a visual distinction between the cement and native bone, and it can also accelerate the hardening process. Certain medical devices incorporate Methylene Blue as a visualization aid. Finally, it is used in construction and soil science to determine the methylene blue value of fine aggregate, an indicator of its clay mineral content. The sheer diversity of these non-medical applications underscores the multifaceted nature of Methylene Blue, stemming from its unique chemical and physical properties [Chain of thought: From industrial applications to educational demonstrations, its unique characteristics make it valuable in diverse fields.].
7. Identify Any Potential Health Benefits of Methylene Blue That Are Not Yet Fully Established or Approved by Regulatory Bodies
7.1 Cognitive Enhancement and Anti-Aging
Emerging research suggests that Methylene Blue may possess potential health benefits beyond its currently approved medical uses, particularly in the areas of cognitive enhancement and anti-aging. Some studies indicate that it may have cognitive-enhancing effects, potentially improving memory and attention span. Animal studies have shown that Methylene Blue can improve age-related memory decline and enhance grip strength and spatial memory in older mice. These effects may be linked to its ability to enhance mitochondrial function in brain cells, which is crucial for energy production and overall cognitive health. Furthermore, Methylene Blue exhibits antioxidant properties that could protect cells, including neurons, against damage from oxidative stress, a process implicated in aging. While these findings from preclinical studies and some early-phase clinical trials are promising, more rigorous and large-scale clinical trials in humans are necessary to definitively establish these benefits and determine safe and effective dosages. It is important to note that over-the-counter Methylene Blue products marketed for cognitive enhancement or anti-aging are not currently regulated by the FDA, and their safety and efficacy have not been fully evaluated .
7.2 Mental Health Support (Beyond Approved Uses)
Research is also exploring the potential of Methylene Blue in providing support for various mental health conditions beyond its historical use in psychosis. Some studies have investigated its use as an adjunct treatment for mood disorders such as depression and bipolar disorder. It is believed that Methylene Blue may influence the levels of certain neurotransmitters in the brain, including serotonin, norepinephrine, and acetylcholine, which play critical roles in mood regulation. Animal models have shown that Methylene Blue exhibits antidepressant-like activity. While these preliminary findings are encouraging, the use of Methylene Blue for mental health support is still considered investigational. Careful consideration of potential drug interactions, particularly with medications that affect serotonin levels, is essential due to Methylene Blue’s properties as a monoamine oxidase inhibitor .
7.3 Lyme Disease and Co-infections
Some practitioners, particularly Lyme Literate Medical Doctors (LLMDs), are exploring the off-label use of Methylene Blue for the treatment of Lyme disease and associated co-infections, such as Bartonella. Anecdotal reports suggest that patients treated with Methylene Blue have experienced improvements in symptoms like fatigue, depression, and cognitive fog, which are commonly associated with these tick-borne illnesses. Laboratory studies have indicated that Methylene Blue possesses antimicrobial properties and may be particularly effective against persister biofilms, a form of bacterial growth that can be difficult to eradicate in Lyme disease and Bartonella infections. However, it is crucial to emphasize that these uses are not yet fully established or approved by regulatory bodies. Rigorous clinical trials in humans are needed to determine the efficacy and safety of Methylene Blue for the treatment of Lyme disease and co-infections.
8. Research the Known Risks, Side Effects, and Contraindications Associated with the Use of Methylene Blue
8.1 Common Side Effects
The use of Methylene Blue is associated with several known side effects, the most common of which is a noticeable bluish-green discoloration of the urine and stool. Some individuals may experience pain in their limbs following intravenous administration. Gastrointestinal disturbances such as nausea, vomiting, diarrhea, and abdominal pain have also been reported. Other common side effects can include dizziness, confusion, and headaches , as well as staining of the mouth or teeth and an altered sense of taste. Some patients may also experience sweating , a burning sensation in the mouth and stomach , restlessness, apprehension, and an unusual taste sensation known as dysgeusia. It is important to note that Methylene Blue can transiently interfere with pulse oximeter readings, potentially leading to an underestimation of the actual oxygen saturation in the blood. Additionally, a decrease in the Bispectral Index (BIS), a measure of brain activity, has been observed following the administration of Methylene Blue during surgical procedures [Chain of thought: Many side effects are relatively mild and related to its properties as a dye and its biochemical actions. However, some can be more significant and require monitoring.].
8.2 Serious Risks and Contraindications
While many side effects are mild, Methylene Blue carries the risk of several serious adverse events and has specific contraindications. One of the most significant risks is the development of serotonin syndrome, a potentially life-threatening condition that can occur when Methylene Blue is used in combination with other drugs that increase serotonin levels in the brain, such as selective serotonin reuptake inhibitors (SSRIs), serotonin-norepinephrine reuptake inhibitors (SNRIs), monoamine oxidase inhibitors (MAOIs), tricyclic antidepressants (TCAs), opioids, and dextromethorphan. Symptoms of serotonin syndrome can include mental status changes, muscle twitching, excessive sweating, shivering, diarrhea, loss of coordination, and fever. Therefore, the concomitant use of Methylene Blue with serotonergic drugs should be avoided. Another serious risk is hemolytic anemia, which is more likely to occur in individuals with glucose-6-phosphate dehydrogenase (G6PD) deficiency. In these patients, Methylene Blue is contraindicated due to the risk of severe hemolysis , which can lead to the formation of Heinz bodies, elevated bilirubin levels, and low haptoglobin. Paradoxically, high doses of Methylene Blue can actually induce methemoglobinemia, the very condition it is used to treat. The administration of Methylene Blue in neonates carries significant risks, including hyperbilirubinemia, respiratory depression, pulmonary edema, phototoxicity, and hemolytic anemia. Methylene Blue is also contraindicated in patients with a known history of hypersensitivity or anaphylactic reactions to it. Its use is contraindicated during pregnancy (FDA pregnancy category X) due to the potential for fetal harm , and it should be avoided by breastfeeding women. Elderly patients with impaired kidney function may require dosage adjustments , and it is contraindicated in cases of severe renal insufficiency. Caution is also advised when using Methylene Blue in patients with hepatic impairment. Beyond serotonergic drugs, Methylene Blue can interact with other medications, including those metabolized by cytochrome P450 enzymes. As a monoamine oxidase inhibitor, it can interact with various substances. It should not be used concurrently with dapsone. Furthermore, if sodium nitrite is used as an antidote for cyanide poisoning, Methylene Blue should not be administered to treat the resulting methemoglobinemia, as this can reduce cyanide binding and increase toxicity .
Table 4: Known Risks, Side Effects, and Contraindications of Methylene Blue
Category
Specific Risk/Side Effect/Contraindication
Relevant Notes/Conditions
Common Side Effects
Bluish-green discoloration of urine and stool
Expected
Common Side Effects
Limb pain following IV administration
Common Side Effects
Nausea, vomiting, diarrhea, abdominal pain
Common Side Effects
Dizziness, confusion, headaches
Common Side Effects
Stained mouth or teeth
Common Side Effects
Altered sense of taste
Common Side Effects
Sweating
Common Side Effects
Burning sensation of the mouth and stomach
Common Side Effects
Restlessness, apprehension, dysgeusia
Common Side Effects
Transiently alters pulse oximeter readings
May underestimate oxygen saturation
Common Side Effects
Fall in Bispectral Index (BIS)
During surgery
Serious Risks
Serotonin Syndrome
With concomitant use of serotonergic drugs and opioids
Oklahoma State Senator Ralph Shortey introduced a bill that would ban “the manufacture or sale of food or products which use aborted human fetuses.”
PepsiCo for working with a company called Senomyx that “has been accused of using proteins derived from human embryonic kidney cells in its research.” Address San Diego, CA 92121-3051
United States Phone+1-858-6468300 Fax 858-4040752 Webwww.senomyx.com
HEK 293 cells But some food companies are using cell lines that were originally derived from human fetuses in order to develop new food products. … The cells, called HEK 293 cells (that stands for human embryonic kidney) were taken from an aborted fetus in the 1970s in the Netherlands. Human cloned DNA in your foods people no joke
The cells, called HEK 293 cells (that stands for human embryonic kidney) were taken from an aborted fetus in the 1970s in the Netherlands.
The Original Unnamed aborted babies cell culture cell line isn’t leading to new abortions but it sure is human DNA.
Origins of the HEK293 Cell Line
HEK293 is a cell line derived from human embryonic kidney cells grown in tissue culture. They are also known, more informally, as HEK cells. This particular line was initiated by the transformation and culturing of normal HEK cells with sheared adenovirus 5 DNA. The transformation resulted in the incorporation of approximately 4.5 kilobases from the viral genome into human chromosome 19 of the HEK cells. The line was cultured by scientist Alex Van der Eb in the early 1970s at his lab at the University of Leiden, Holland. The transformation was executed by Frank Graham, another scientist Van der Eb’s lab who invented the calcium phosphate method for transfecting cells. The source of the cells was a healthy aborted fetus of unknown parenthood. The name HEK293 is thusly named because it was Frank Graham’s 293rd experiment.
The type of kidney cell that the HEK293 cell line came arose from is unknown and it is difficult to conclusively characterize the cells post-transformation since adenovirus 5 could have significantly disrupted cell morphology and expression. Also, embryonic kidneys are a heterogeneous mix of almost all the types of cells present in the body. In fact, it has been speculated by independent researchers, including Van der Eb himself, that the cells may be neuronal in origin. Although theoretically possible, most cells derived from an embryonic kidney would be endothelial, epithelial or fibroblast cells. Neuronal origin is suspected due to the presence of mRNA and gene products typically found in neurons.
Today, HEK293 cells are frequently used in cell biology and biotechnology, second only to HeLa, the first human cell line. Around establishment of HeLa in 1951, scientists were reluctant to accept and use human cell lines out of concern for an oncogenic agent in them. This concern, along with the known ability of animal cell lines to grow rapidly and yield a high amount of proteins, gave scientists reason to favor animal cell lines over human cell lines when producing recombinant proteins. However, advances in technology since then have allowed for an increase in human cell line use. One advantage of human cell lines is that they are able to produce proteins most similar to those that humans naturally synthesize. Now there are approved recombinant biotherapeutic products produced from HEK293 and other human cell lines.
HEK293 and its derivatives are used in a wide range of experiments, including signal transduction and protein interaction studies, rapid small-scale protein production, and biopharmaceutical production. HEK293 cells easily grow in suspension serum-free culture, reproduce rapidly, and produce high levels of protein, which explains why they have been widely used to produce research-grade proteins for a number of years.
“We’re helping companies clean up their labels,” said Senomyx’s chief executive, Kent Snyder.
Senomyx, based in San Diego, uses many of the same research techniques that biotechnology companies apply in devising new drugs. Executives say that a taste receptor or family of receptors on the tongue or in the mouth are responsible for recognizing a taste. Using the human genome sequence, the company says, it has identified hundreds of those taste receptors. Its chemical compounds activate the receptors in a way that accentuates the taste of sugar or salt. It is still experimenting to determine the most potent compounds, its chief scientist, Mark Zoller, said.
But Senomyx maintains that its new products are safe because they will be used in tiny quantities.
Kraft, Nestlé, Coca-Cola and Campbell Soup have contracted with Senomyx for exclusive rights to use the ingredients in certain types of food and beverages, although the companies declined to identify those categories.
Elise Wang, an analyst at Smith Barney, said that Kraft was planning to use Senomyx’s sweet flavoring to reduce the sugar in powdered beverages like Kool-Aid by one-third. Campbell Soup, she said, is looking at cutting sodium levels by a third with the salt flavoring.
Cost Effective
Since Cytofect™ Transfection Reagents require no expensive instruments to use, high efficiency transfection of primary cells becomes much less costly.
With no upfront investment in electroporation devices, your lab can save its precious research budget to focus on downstream assays instead.
Senomyx is an Americanbiotechnology company working toward developing additives to amplify certain flavors and smells in foods. The company claims to have essentially “reverse engineered” the receptors in humans that react for taste and aroma, and that they are capitalizing on these discoveries to produce chemicals that will make food taste better. On 17 Sept 2018, Firmenich completed the acquisition of Senomyx. [1]
Senomyx develops patented flavor enhancers by using “proprietary taste receptor-based assay systems”, which have been previously expressed in human cell culture, in HEK293 cells.[2]
HEK293 cells are a cell line widely used in biological and medical research, immortalised through a genetic modification removed from the original human embryonic kidney cells taken from a healthy, electively aborted human fetus in the early 1970s.[3] The receptors in the assay are used to identify flavours; they are not used as flavours themselves. No human taste receptors are used as ingredients in any flavourings. Using information from the human genome sequence, Senomyx has identified hundreds of taste receptors and currently owns 113 patents on their discoveries. Senomyx collaborates with seven of the world’s largest food companies to further their research and to fund development of their technology.
Cell Applications, Inc
5820 Oberlin Drive, Suite 101
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types, complimented by optimized products to serve life science R&D … media like DMEM and RPMI, and immortalized cell lines (HeLa, HEK 293 … testing and certification, so the procedures and product remains consistent. …
I. TERMS OF USE Cell Applications, Inc. (“CAI”) will sell products … of the CAI’s products (such as to collect a debt, resolve a dispute … by delays in receiving orders. VI. PRODUCT USE AND RECOMMENDATIONS All …
Srihirun, S., Park, J. W., Teng, R., Sawaengdee, W., Piknova, B., & Schechter, A. N. (2019). Nitrate uptake and metabolism in human skeletal muscle cell cultures. Nitric Oxide.
Human Aortic Smooth Muscle Cells: HAOSMC
Lu, Y., Sun, X., Peng, L., Jiang, W., Li, W., Yuan, H., & Cai, J. (2019). Angiotensin II-Induced vascular remodeling and hypertension involves cathepsin L/V-MEK/ERK mediated mechanism. International Journal of Cardiology.
Human Dermal Fibroblasts: HDF
Chaudhuri, R. K., Meyer, T., Premi, S., & Brash, D. Acetyl Zingerone (2019): An efficacious multifunctional ingredient for continued protection against on‐going DNA damage in melanocytes after sun exposure ends. International Journal of Cosmetic Science.
Human Coronary Artery Endothelial Cells: HCAEC
Gamon, L. F., Dieterich, S., Ignasiak, M. T., Schrameyer, V., & Davies, M. J. (2019). Iodide modulates protein damage induced by the inflammation-associated heme enzyme myeloperoxidase. Redox Biology, 101331.
Rat Dermal Fibroblasts: RDF
Palungwachira, P., Tancharoen, S., Phruksaniyom, C., Klungsaeng, S., Srichan, R., Kikuchi, K., & Nararatwanchai, T. (2019). Antioxidant and Anti-Inflammatory Properties of Anthocyanins Extracted from Oryza sativa L. in Primary Dermal Fibroblasts. Oxidative Medicine and Cellular Longevity, 2019.
Human Dermal Fibroblasts: HDF
Wang, X., Hong, H., & Wu, J. (2019). Hen collagen hydrolysate alleviates UVA-induced damage in human dermal fibroblasts. Journal of Functional Foods, 63, 103574.
Human Epidermal Keratinocytes: HEK
Chaudhuri, R. K., Meyer, T., Premi, S., & Brash, D. Acetyl Zingerone (2019): An efficacious multifunctional ingredient for continued protection against on‐going DNA damage in melanocytes after sun exposure ends. International Journal of Cosmetic Science.
Human Umbilical Vein Endothelial Cells: HUVEC
Matsunuma, S., Handa, S., Kamei, D., Yamamoto, H., Okuyama, K., & Kato, Y. (2019). Oxaliplatin induces prostaglandin E2 release in vascular endothelial cells. Cancer Chemotherapy and Pharmacology, 1-6.
Human Pulmonary Artery Endothelial Cells: HPAEC
Blais-Lecours, P., Laouafa, S., Arias-Reyes, C., Santos, W. L., Joseph, V., Burgess, J. K., … & Marsolais, D. (2019). Metabolic adaptation of airway smooth muscle cells to a SPHK2 substrate precedes cytostasis. American Journal of Respiratory Cell and Molecular Biology,
Bovine Aortic Endothelial Cells: BAOEC
Ogata, F., Nakamura, T., Nakajima, M., Toda, M., Otani, M., & Kawasaki, N. (2019). PO43− adsorption in a complex solution by nickel–cobalt hydroxide, and its cytotoxicity on bovine aortic endothelial cells. Journal of Environmental Chemical Engineering.
MesoEndo Cell Growth Medium
Detsika, M. G., Myrtsi, E. D., Koulocheri, S. D., Haroutounian, S. A., Lianos, E. A., & Roussos, C. (2019). Induction of decay accelerating factor and membrane cofactor protein by resveratrol attenuates complement deposition in human coronary artery endothelial cells. Biochemistry and Biophysics Reports, 19, 100652.
Rat Pulmonary Artery Smooth Muscle Cells: RPASMC
Suzuki, Y. J., Marcocci, L., Shimomura, T., Tatenaka, Y., Ohuchi, Y., & Brelidze, T. I. (2019). Protein Redox State Monitoring Studies of Thiol Reactivity. Antioxidants, 8 (5), 143.
Human Coronary Artery Endothelial Cells: HCAEC
Lorentzen, L. G., Chuang, C. Y., Rogowska-Wrzesinska, A., & Davies, M. J. (2019). Identification and quantification of sites of nitration and oxidation in the key matrix protein laminin and the structural consequences of these modifications. Redox Biology, 101226.
Human Liver, Spleen, Kidney and Testes RNA
Swystun, L. L., Ogiwara, K., Lai, J. D., Ojala, J. R., Rawley, O., Lassalle, F., … & Tryggvason, K. (2019). The scavenger receptor SCARA 5 is an endocytic receptor for von Willebrand factor expressed by littoral cells in the human spleen. Journal of Thrombosis and Haemostasis.
Human Umbilical Vein Endothelial Cells: HUVEC
Brines, M. and Cerami, A., (2019). TISSUE PROTECTIVE PEPTIDES FOR PREVENTING AND TREATING DISEASES AND DISORDERS ASSOCIATED WITH TISSUE DAMAGE. U.S. Patent Application 16/096,247.
Human Dermal Fibroblasts: HDF
Yang, H., Sun, J., Chen, H., Wang, F., Li, Y., Wang, H., & Qu, T. (2019). Mesenchymal stem cells from bone marrow attenuated the chronic morphine-induced cAMP accumulation in vitro. Neuroscience letters, 698, 76-80.
Human EpiVita Serum-Free Growth Medium
Lin, E. S., Chang, W. A., Chen, Y. Y., Wu, L. Y., Chen, Y. J., & Kuo, P. L. (2019). Deduction of Novel Genes Potentially Involved in Keratinocytes of Type 2 Diabetes Using Next-Generation Sequencing and Bioinformatics Approaches. Journal of clinical medicine, 8(1), 73.
Human Carotid Artery Smooth Muscle Cells: HCtASMC
Aldi, S., Eriksson, L., Kronqvist, M., Lengquist, M., Löfling, M., Folkersen, L…& Österholm, C. (2019). Dual roles of heparanase in human carotid plaque calcification. Atherosclerosis.
Human Umbilical Vein Endothelial Cells: HUVEC
Swaminathan, S., Hamid, Q., Sun, W., & Clyne, A. M. (2019). Bioprinting of 3D breast epithelial spheroids for human cancer models. Biofabrication.
MesoEndo Cell Growth Medium
Pott, G. B., Tsurudome, M., Proctor, L. L., & Goalstone, M. L. (2019). CIGARETTE SMOKE EXTRACT, KALLIKREIN-6 AND APROTININ REGULATE PRODUCTION OF SOLUBLE VCAM-1 AND ICAM-1 IN HUMAN CAROTID ENDOTHELIAL CELLS.
Human Epidermal Keratinocytes: HEK
Yamakami, Y., Morino, K., Takauji, Y., Kasukabe, R., Miki, K., Hossain, M. N., … & Fujii, M. (2019). Extract of Emblica officinalis enhances the growth of human keratinocytes in culture. Journal of integrative medicine.
Human Bladder Epithelial Cells: HBlEpC
Kim, D., Ahn, B. N., Kim, Y., Hur, D. Y., Yang, J. W., Park, G. B., … & Kim, M. K. (2019). High Glucose with Insulin Induces Cell Cycle Progression and Activation of Oncogenic Signaling of Bladder Epithelial Cells Cotreated with Metformin and Pioglitazone. Journal of diabetes research, 2019.
Human Carotid Artery Endothelial Cells: HCtAEC
Pott, G. B., Tsurudome, M., Proctor, L. L., & Goalstone, M. L. (2019). CIGARETTE SMOKE EXTRACT, KALLIKREIN-6 AND APROTININ REGULATE PRODUCTION OF SOLUBLE VCAM-1 AND ICAM-1 IN HUMAN CAROTID ENDOTHELIAL CELLS.
Human Dermal Fibroblasts: HDF
Desai, D., Lauver, M. D., Cruz, L., Jin, G., Ferguson, K., Roper, B., … & Buchkovich, N. J. (2019). Inhibition of Diverse Opportunistic Viruses by Structurally Optimized Retrograde Trafficking Inhibitors. Bioorganic & Medicinal Chemistry.
Human Mammary Epithelial Cells: HMEpC
Fukui, T., Soda, K., Takao, K., & Rikiyama, T. (2019). Extracellular spermine activates DNA methyltransferase 3A and 3B. International journal of molecular sciences, 20(5), 1254.
Rat Aortic Endothelial Cells: RAOEC
Naik, J. S., & Walker, B. R. (2018). Endothelial-dependent dilation following chronic hypoxia involves TRPV4-mediated activation of endothelial BK channels. Pflügers Archiv-European Journal of Physiology, 470(4), 633-648.
2018
Human Chondrocytes
Chen, Y.J., Chang, W.A., Wu, L.Y., Hsu, Y.L., Chen, C.H. and Kuo, P.L., 2018. Systematic Analysis of Transcriptomic Profile of Chondrocytes in Osteoarthritic Knee Using Next-Generation Sequencing and Bioinformatics. Journal of Clinical Medicine, 7(12), p.535.
Bovine Aortic Endothelial Cells: BAOEC
Takahashi, A., Takahashi, M., Fujie, T., Hara, T., Yoshida, E., Yamamoto, C. and Kaji, T., 2018. A zinc complex that suppresses the expression of a reactive sulfur species-producing enzyme, cystathionine γ-lyase, in cultured vascular endothelial cells. Fundamental Toxicological Sciences, 5(6), pp.181-184.
Human Dermal Fibroblasts: HDF
Yu, C., Ma, X., Zhu, W., Wang, P., Miller, K.L., Stupin, J., Koroleva-Maharajh, A., Hairabedian, A. and Chen, S., 2018. Scanningless and continuous 3D bioprinting of human tissues with decellularized extracellular matrix. Biomaterials.
Human Umbilical Vein Endothelial Cells: HUVEC
Tan, Z. B., Fan, H. J., Wu, Y. T., Xie, L. P., Bi, Y. M., Xu, H. L., … & Zhou, Y. C. (2018). Rheum palmatum extract exerts anti-hepatocellular carcinoma effects by inhibiting signal transducer and activator of transcription 3 signaling. Journal of Ethnopharmacology.
Skeletal Muscle Growth Medium
Patton, J. B., Bennuru, S., Eberhard, M. L., Hess, J. A., Torigian, A., Lustigman, S., … & Abraham, D. (2018). Development of Onchocerca volvulus in humanized NSG mice and detection of parasite biomarkers in urine and serum. PLOS Neglected Tropical Diseases, 12(12), e0006977.
Human Chondrocytes
Tsumaki, N. and Yamashita, A., Kyoto University, 2018. Prophylactic and therapeutic agents for fgfr3 diseases and screening method for the same. U.S. Patent Application 16/059,462.
Human Dermal Fibroblasts: HDF
Playne, R., Jones, K. S., & Connor, B. (2018). Generation of dopamine neuronal-like cells from induced neural precursors derived from adult human cells by non-viral expression of lineage factors. J Stem Cells Regen Med.
Human Dermal Fibroblasts: HDF
Ikeda, K., Uchida, N., Nishimura, T., White, J., Martin, R.M., Nakauchi, H., Sebastiano, V., Weinberg, K.I. and Porteus, M.H., (2018). Efficient scarless genome editing in human pluripotent stem cells. Nature methods, 15(12), p.1045.
Endothelial Cell Growth Medium Leonard, J.N., Stranford, D.M. and Passineau, M.J., Northwestern University, (2018). Deliverable extracellular vesicles incorporating cell membrane transport proteins. U.S. Patent Application 15/975,222.
Human Peripheral Blood B Cells: HPBB
Marin, E.H., Paek, H., Li, M., Ban, Y., Karaga, M.K., Shashidharamurthy, R. and Wang, X., 2018. Caffeic acid phenethyl ester exerts apoptotic and oxidative stress on human multiple myeloma cells. Investigational new drugs, pp.1-12.
Human Adipocyte Differentiation Medium
Bagher, Z., Kamrava, S. K., Alizadeh, R., Farhadi, M., Absalan, M., Falah, M. & Komeili, A. (2018). Differentiation of Neural Crest Stem Cells From Nasal Mucosa into Motor Neuron-Like Cells. Journal of Chemical Neuroanatomy.
MCDB 105 Medium
Starbuck, K., Al-Alem, L., Eavarone, D. A., Hernandez, S. F., Bellio, C., Prendergast, J. M., & Behrens, J. (2018). Treatment of ovarian cancer by targeting the tumor stem cell-associated carbohydrate antigen, Sialyl-Thomsen-nouveau. Oncotarget, 9(33), 23289.
Bovine Pulmonary Artery Endothelial cells: BPAEC
Rowan, S. C., Rochfort, K. D., Piouceau, L., Cummins, P. M., O’Rourke, M., & McLoughlin, P. (2018). Pulmonary endothelial permeability and tissue fluid balance depend on the viscosity of the perfusion solution. American Journal of Physiology-Lung Cellular and Molecular Physiology.
Human Dermal Fibroblasts: HDF
Chaudhuri, R.K., Sytheon Ltd, 2018. Skin enhancing compositions and methods. U.S. Patent Application 15/798,804.
Human Preadipocytes: HPAd
Matsubara, Yumiko, Takeru Zama, Yasuo Ikeda, Yukako Uruga, Toshio Suda, and Sahoko Matsuoka. “Method for producing megakaryocytes, platelets and/or thrombopoietin using mesenchymal cells.” U.S. Patent Application 15/815,069.
Human Aortic Smooth Muscle Cells: HAOSMC
van Engeland, N. C., Pollet, A. M., den Toonder, J. M., Bouten, C. V., Stassen, O. M., & Sahlgren, C. M. (2018). A biomimetic microfluidic model to study signalling between endothelial and vascular smooth muscle cells under hemodynamic conditions. Lab on a Chip.
Canine Osteoblasts: CnOb
Scott, M.C., Sarver, A.L., Modiano, J.F., Subramanian, S., Largaespada, D.A. and Spector, L.G., University of Minnesota, 2018. Tumor Analytical Methods. U.S. Patent Application 15/783,352.
Human Dermal Fibroblasts: HDF
Yoshida, Shunsuke, Mitsuru Inamura, Tohru Tanaka, Hiroyuki Ishikawa, and Hidenori Ito. “Stem cell removing method, differentiated cell protective method, and culture medium composition.” U.S. Patent Application 15/565,422.
Human Chondrocytes: HC
Li, A., Wei, Y., Hung, C., & Vunjak-Novakovic, G. (2018). Chondrogenic properties of collagen type XI, a component of cartilage extracellular matrix. Biomaterials.
Human Coronary Artery Endothelial Cells: HCAEC
Xu, S., Xu, Y., Yin, M., Zhang, S., Liu, P., Koroleva, M.,..& Jin, Z. G. (2018). Flow-dependent epigenetic regulation of IGFBP5 expression by H3K27me3 contributes to endothelial anti-inflammatory effects. Theranostics, 8(11), 3007-3021.
Human MesoEndo Endothelial Cell Media
Xu, S., Xu, Y., Yin, M., Zhang, S., Liu, P., Koroleva, M.,..& Jin, Z. G. (2018). Flow-dependent epigenetic regulation of IGFBP5 expression by H3K27me3 contributes to endothelial anti-inflammatory effects. Theranostics, 8(11), 3007-3021.
Rat Aortic Smooth Muscle Cells: RAOSMC
Park, H. S., Han, J. H., Jung, S. H., Lee, D. H., Heo, K. S., & Myung, C. S. (2018). Anti-apoptotic effects of autophagy via ROS regulation in microtubule-targeted and PDGF-stimulated vascular smooth muscle cells. The Korean Journal of Physiology & Pharmacology, 22(3), 349-360.
Human Dermal Fibroblasts: HDF
Kikkawa, Y., Enomoto-Okawa, Y., Fujiyama, A., Fukuhara, T., Harashima, N., Sugawara, Y., … & Ito, Y. (2018). Internalization of CD239 highly expressed in breast cancer cells: a potential antigen for antibody-drug conjugates. Scientific reports, 8.
Human Pulmonary Artery Smooth Muscle Cells: HPASMC
Wilson, J. L., Warburton, R., Taylor, L., Toksoz, D., Hill, N., & Polgar, P. (2018). Unraveling endothelin-1 induced hypercontractility of human pulmonary artery smooth muscle cells from patients with pulmonary arterial hypertension. PloS one, 13(4), e0195780.
Human Dermal Fibroblasts: HDF
Ito, Tomohisa, Takashi Ando, Miki Suzuki-Karasaki, Tomohiko Tokunaga, Yukihiro Yoshida, Toyoko Ochiai, Yasuaki Tokuhashi, and Yoshihiro Suzuki-Karasaki. “Cold PSM, but not TRAIL, triggers autophagic cell death: A therapeutic advantage of PSM over TRAIL.” International Journal of Oncology.
Human Carotid Artery Endothelial Cells: HCtAEC
Hoh, B. L., Rojas, K., Lin, L., Fazal, H. Z., Hourani, S., Nowicki, K. W., … & Hosaka, K. (2018). Estrogen Deficiency Promotes Cerebral Aneurysm Rupture by Upregulation of Th17 Cells and Interleukin‐17A Which Downregulates E‐Cadherin. Journal of the American Heart Association, 7(8), e008863.
Sakima, M., Hayashi, H., Al Mamun, A., & Sato, M. (2018). VEGFR-3 signaling is regulated by a G-protein activator, activator of G-protein signaling 8, in lymphatic endothelial cells. Experimental cell research.
Human Dermal Fibroblasts: HDF
Kang, L., Liu, X., Yue, Z., Chen, Z., Baker, C., Winberg, P. C., & Wallace, G. G. (2018). Fabrication and In Vitro Characterization of Electrochemically Compacted Collagen/Sulfated Xylorhamnoglycuronan Matrix for Wound Healing Applications. Polymers, 10(4), 415.
Human Chondrocyte Media
Barrett, Carolyn, and Yaling Shi. “Cartilage mosaic compositions and methods.” U.S. Patent Application 15/608,679.
Human Dermal Fibroblasts: HDF
Esparza, Y., Bandara, N., Ullah, A., & Wu, J. (2018). Hydrogels from feather keratin show higher viscoelastic properties and cell proliferation than those from hair and wool keratins. Materials Science and Engineering: C.
Human Aortic Smooth Muscle Cells: HAOSMC
Cardenas, C. L. L., Kessinger, C. W., Cheng, Y., MacDonald, C., MacGillivray, T., Ghoshhajra, B., … & Kaminski, N. (2018). An HDAC9-MALAT1-BRG1 complex mediates smooth muscle dysfunction in thoracic aortic aneurysm. Nature Communications, 9(1), 1009.
Human Epidermal Keratinocytes: HEK
Takahashi, A., Loo, T. M., Okada, R., Kamachi, F., Watanabe, Y., Wakita, M., & Ohtani, N. (2018). Downregulation of cytoplasmic DNases is implicated in cytoplasmic DNA accumulation and SASP in senescent cells. Nature Communications, 9(1), 1249.
Bovine Aortic Endothelial Cells: BAOEC
Zhao, X., Hui, D. S., Lee, R., & Edwards, J. L. (2018). Ratiometric quantitation of thiol metabolites using non-isotopic mass tags. Analytica Chimica Acta.
Human Endothelial Cell Growth Medium
Passineau, M.J., Murali, S., Benza, R.L. and Pollett, J.B., Allegheny-Singer Research Institute, 2018. ISOLATION OF PULMONARY ARTERIAL ENDOTHELIAL CELLS FROM PATIENTS WITH PULMONARY VASCULAR DISEASE AND USES THEREOF. U.S. Patent Application 15/806,751.
DiI-Ac-LDL Kit
Lian, W., Hu, X., Shi, R., Han, S., Cao, C., Wang, K., & Li, M. (2018). MiR-31 regulates the function of diabetic endothelial progenitor cells by targeting Satb2. Acta biochimica et biophysica Sinica.
Human Hair Follicle Dermal Papilla Cells: HFDPC
Lahiji SF, Seo SH, Kim S, Dangol M, Shim J, Li CG, Ma Y, Lee C, Kang G, Yang H, Choi KY. (2018). Transcutaneous implantation of valproic acid-encapsulated dissolving microneedles induces hair regrowth. Biomaterials.
Bovine Aortic Endothelial Cells: BAOEC
Zhao, X., Hui, D. S., Lee, R., & Edwards, J. L. (2018). Ratiometric quantitation of thiol metabolites using non-isotopic mass tags. Analytica Chimica Acta.
Human Aortic Smooth Muscle Cells: HAOSMC
Cardenas, C. L. L., Kessinger, C. W., MacDonald, C., Jassar, A. S., Isselbacher, E. M., Jaffer, F. A., & Lindsay, M. E. (2018). Inhibition of the methyltranferase EZH2 improves aortic performance in experimental thoracic aortic aneurysm. JCI insight, 3(5).
Endothelial Cell Growth Medium
CD Nichols, B YU (2018). LOW DOSAGE SEROTONIN 5-HT2A RECEPTOR AGONIST TO SUPPRESS INFLAMMATION. US Patent App. 15/478,437.
Rat Brain Microvascular Endothelial Cells: RBMVEC
Brailoiu, E., Barlow, C. L., Ramirez, S. H., Abood, M. E., & Brailoiu, G. C. (2018). Effects of Platelet-Activating Factor on brain microvascular endothelial cells. Neuroscience.
Human Carotid Artery Smooth Muscle Cells: HCtASMC
Han, X., Sakamoto, N., Tomita, N., Meng, H., Sato, M., & Ohta, M. (2017). Influence of shear stress on phenotype and MMP production of smooth muscle cells in a co-culture model. Journal of Biorheology, 31(2), 50-56.
Human Fibroblast-Like Synoviocytes: HFLS
Yu, R., Li, C., Sun, L., Jian, L., Ma, Z., Zhao, J., & Liu, X. (2018). Hypoxia induces production of citrullinated proteins in human fibroblast‐like synoviocytes through regulating HIF1α. Scandinavian journal of immunology.
Human Cardiac Fibroblasts: HCF
John, C.M., Meenakshi, G.A.U.R., Matthew, L. and Wang, X., MANDALMED Inc, 2018. Methods and compositions for preventing and treating damage to the heart. U.S. Patent Application 15/666,456.
Rat Smooth Muscle Cell Media
Chinnappan, M., Mohan, A., Agarwal, S., Dalvi, P., & Dhillon, N. K. (2018). Network of MicroRNAs Mediate Translational Repression of Bone Morphogenetic Protein Receptor‐2: Involvement in HIV‐Associated Pulmonary Vascular Remodeling. Journal of the American Heart Association, 7(5), e008472.
Human Smooth Muscle Cell Growth Medium
Cardenas, C. L. L., Kessinger, C. W., MacDonald, C., Jassar, A. S., Isselbacher, E. M., Jaffer, F. A., & Lindsay, M. E. (2018). Inhibition of the methyltranferase EZH2 improves aortic performance in experimental thoracic aortic aneurysm. JCI insight, 3(5).
Human Fibroblast-Like Synoviocytes: HFLS
Rosa, I., Marini, M., Guasti, D., Ibba-Manneschi, L., & Manetti, M. (2018). Morphological evidence of telocytes in human synovium. Scientific reports, 8(1), 3581.
Human Carotid Artery Endothelial Cells: HCtAEC
Han, X., Sakamoto, N., Tomita, N., Meng, H., Sato, M., & Ohta, M. (2017). Influence of shear stress on phenotype and MMP production of smooth muscle cells in a co-culture model. Journal of Biorheology, 31(2), 50-56.
Human Fibroblast-Like Synoviocytes: Rheumatoid Arthritis: HFLS-RA
Hagihara, M., Shimizu, M. and Wada, Y., Ube Industries Ltd, 2018. Method of producing substance. U.S. Patent Application 15/545,624.
Bovine Aortic Endothelial Cells: BAOEC
Uhl, C. G., Gao, Y., Zhou, S., & Liu, Y. (2018). The shape effect on polymer nanoparticle transport in a blood vessel. RSC Advances, 8(15), 8089-8100.
Human Umbilical Vein Endothelial Cells: HUVEC
Sasahira, T., Nishiguchi, Y., Kurihara-Shimomura, M., Nakashima, C., Kuniyasu, H., & Kirita, T. (2018). NIPA-like domain containing 1 is a novel tumor-promoting factor in oral squamous cell carcinoma. Journal of cancer research and clinical oncology, 1-8.
Human Fibroblast-Like Synoviocytes: Rheumatoid Arthritis: HFLS-RARhys, H. I., Dell’Accio, F., Pitzalis, C., Moore, A., Norling, L. V., & Perretti, M. (2018). Neutrophil Microvesicles from Healthy Control and Rheumatoid Arthritis Patients Prevent the Inflammatory Activation of Macrophages. EBioMedicine.
Rabbit Aortic Smooth Muscle Cells: RbAOSMC
Honda, K., Matoba, T., Antoku, Y., Koga, J. I., Ichi, I., Nakano, K., & Egashira, K. (2018). Lipid-Lowering Therapy With Ezetimibe Decreases Spontaneous Atherothrombotic Occlusions in a Rabbit Model of Plaque ErosionHighlights: A Role of Serum Oxysterols. Arteriosclerosis, thrombosis, and vascular biology, 38(4), 757-771.
Human Dermal Fibroblasts: HDF
Tokunaga, T., Ando, T., Suzuki-Karasaki, M., Ito, T., Onoe-Takahashi, A., Ochiai, T., Soma, M. and Suzuki-Karasaki, Y., 2018. Plasma-stimulated medium kills TRAIL-resistant human malignant cells by promoting caspase-independent cell death via membrane potential and calcium dynamics modulation. International journal of oncology, 52(3), pp.697-708.
Human Coronary Artery Endothelial Cells RNA
Baggio, L. L., Yusta, B., Mulvihill, E. E., Cao, X., Streutker, C. J., Butany, J., & Drucker, D. J. (2018). GLP-1 receptor expression within the human heart. Endocrinology, 159(4), 1570-1584.
Grunlan, M.A., Cote, G.L., Abraham, A.A., Fei, R. and Locke, A.K., Texas A&M University System, 2018. Self-Cleaning Membrane for Medical Devices. U.S. Patent Application 15/545,811.
Bovine Aortic Smooth Muscle Cells: BAOSMC
Tsukagoshi, T., Nguyen, T. V., Shoji, K. H., Takahashi, H., Matsumoto, K., & Shimoyama, I. (2018). Cellular dynamics of bovine aortic smooth muscle cells measured using MEMS force sensors. Journal of Physics D: Applied Physics, 51(14), 145401.
Rat Fibroblast Growth Medium
Grunlan, M.A., Cote, G.L., Abraham, A.A., Fei, R. and Locke, A.K., Texas A&M University System, 2018. Self-Cleaning Membrane for Medical Devices. U.S. Patent Application 15/545,811.
Human Umbilical Vein Endothelial Cells: HUVEC
Gaston, B.M., Straub, A.C., Isakson, B.E. and Columbus, L., University of Virginia Licensing and Ventures Group, 2018. Compositions and methods for regulating arterial tone. U.S. Patent Application 15/643,633.
Rat Aortic Endothelial Cells: RAOEC
Naik, J.S. and Walker, B.R., 2018. Endothelial-dependent dilation following chronic hypoxia involves TRPV4-mediated activation of endothelial BK channels. Pflügers Archiv-European Journal of Physiology, pp.1-16.
Human Fibroblast-Like Synoviocytes: HFLS
Hagihara, M., Shimizu, M. and Wada, Y., Ube Industries Ltd, 2018. Method of producing substance. U.S. Patent Application 15/545,624.
Human Preadipocytes: HPAd
Oishi, T., Sakata, A., Shishido, M. and Hirakawa, S., A serum protein, an unexpected player inducing the skin sagging, and a proposed measure for improving the facial sagging.
Human Adipocyte Differentiation Medium Oishi, T., Sakata, A., Shishido, M. and Hirakawa, S., A serum protein, an unexpected player inducing the skin sagging, and a proposed measure for improving the facial sagging.
Human Umbilical Vein Smooth Muscle Cells: HUVSMC
Gaston, B.M., Straub, A.C., Isakson, B.E. and Columbus, L., University of Virginia Licensing and Ventures Group, 2018. Compositions and methods for regulating arterial tone. U.S. Patent Application 15/643,633.
Rat Aortic Endothelial Cells: RAOEC
Iba, T., Hirota, T., Sato, K. and Nagaoka, I., 2018. Protective effect of a newly developed fucose-deficient recombinant antithrombin against histone-induced endothelial damage. International Journal of Hematology, pp.1-7.
Human Dermal Fibroblasts: HDF
Ito, N., Katoh, K., Kushige, H., Saito, Y., Umemoto, T., Matsuzaki, Y., Kiyonari, H., Kobayashi, D., Soga, M., Era, T. and Araki, N., 2018. Ribosome Incorporation into Somatic Cells Promotes Lineage Transdifferentiation towards Multipotency. Scientific reports, 8(1), p.1634.
Human dermal fibroblast growth medium
Ito, N., Katoh, K., Kushige, H., Saito, Y., Umemoto, T., Matsuzaki, Y., Kiyonari, H., Kobayashi, D., Soga, M., Era, T. and Araki, N., 2018. Ribosome Incorporation into Somatic Cells Promotes Lineage Transdifferentiation towards Multipotency. Scientific reports, 8(1), p.1634.
Human Dermal Fibroblasts: HDF
Martin, R., Ikeda, K., Uchida, N., Cromer, M.K., Nishimura, T., Dever, D.P., Camarena, J., Bak, R., Lausten, A., Jakobsen, M.R. and Wiebking, V., 2018. Selection-free, high frequency genome editing by homologous recombination of human pluripotent stem cells using Cas9 RNP and AAV6. bioRxiv, p.252163.
DiI-Ac-LDL Kit
Iba, T., Hirota, T., Sato, K. and Nagaoka, I., 2018. Protective effect of a newly developed fucose-deficient recombinant antithrombin against histone-induced endothelial damage. International Journal of Hematology, pp.1-7.
Rat cardiac fibroblasts
Fan, Z., Xu, Z., Niu, H., Gao, N., Guan, Y., Li, C., Dang, Y., Cui, X., Liu, X.L., Duan, Y. and Li, H., 2018. An
Injectable Oxygen Release System to Augment Cell Survival and Promote Cardiac Repair Following Myocardial Infarction. Scientific Reports, 8(1), p.1371.
Ishida, K., Xu, H., Sasakawa, N., Lung, M.S.Y., Kudryashev, J.A., Gee, P. and Hotta, A., 2018. Site-specific randomization of the endogenous genome by a regulatable CRISPR-Cas9 piggyBac system in human cells. Scientific reports, 8(1), p.310.
Human Coronary Artery Endothelial Cells: HCAEC
Hwang, H.V., Tran, D.T., Rebuffatti, M.N., Li, C.S. and Knowlton, A.A., 2018. Investigation of quercetin and hyperoside as senolytics in adult human endothelial cells. PloS one, 13(1), p.e0190374.
Human Epidermal Keratinocytes: HEK
Qiao, M., Li, R., Zhao, X., Yan, J. and Sun, Q., 2018. Up-regulated lncRNA-MSX2P1 promotes the growth of IL-22-stimulated keratinocytes by inhibiting miR-6731-5p and activating S100A7. Experimental cell research.
2017
Human Umbilical Vein Endothelial Cells: HUVEC
Izzicupo, P., D’Amico, M.A., Di Blasio, A., Napolitano, G., Nakamura, F.Y., Di Baldassarre, A. and Ghinassi, B., 2017. Aerobic Training Improves Angiogenic Potential Independently of VEGF Modifications in Postmenopausal Women. Frontiers in Endocrinology, 8, p.363.
Human Dermal Fibroblasts: HDF
Ohta, K. and Ito, N., NATIONAL UNIVERSITY CORPORATION KUMAMOTO UNIVERSITY, 2017. METHOD FOR INDUCING CELL REPROGRAMMING, AND METHOD FOR PRODUCING PLURIPOTENT CELLS. U.S. Patent Application 15/310,189.
Human Pulmonary Artery Smooth Muscle Cells: HPASMC
Nadeau, V., Potus, F., BOUCHERAT, O., Paradis, R., Tremblay, E., Iglarz, M., PAULIN, R., Bonnet, S. and PROVENCHER, S., 2017. Dual eta/etb blockade with macitentan improves both vascular remodelling and angiogenesis in pulmonary arterial hypertension. Pulmonary Circulation, p.2045893217741429.
Bovine Pulmonary Artery Endothelial Cells: BPAEC
Frawley, Kristin L., Andrea A. Cronican, Linda Lorraine Pearce, and Jim Peterson., 2017. Sulfide Toxicity and Its Modulation by Nitric Oxide in Bovine Pulmonary Artery Endothelial Cells. Chemical Research in Toxicology (2017).
Classical Cell Media: MCDB-105
He, S., Deng, Y., Liao, Y., Li, X., Liu, J. and Yao, S., 2017. CREB5 promotes tumor cell invasion and correlates with poor prognosis in epithelial ovarian cancer. Oncology Letters, 14(6), pp.8156-8161.
Bovine Brain Endothelial Cell Growth Medium
Duck, K.A., Simpson, I.A. and Connor, J.R., 2017. Regulatory mechanisms for iron transport across the blood-brain barrier. Biochemical and Biophysical Research Communications.
Human Osteoblast Growth Medium
Chen, Y.J., Chang, W.A., Hsu, Y.L., Chen, C.H. and Kuo, P.L., 2017. Deduction of Novel Genes Potentially Involved in Osteoblasts of Rheumatoid Arthritis Using Next-Generation Sequencing and Bioinformatic Approaches. International Journal of Molecular Sciences, 18(11), p.2396.
Bovine Insulin
Buckner, S., Pruitt, A., Thomas, C., Amin, M., Miller, L., Wiley, F. and Sabbatini, M.E., 2017. Di-N-octylphthalate acts as a proliferative agent in murine cell hepatocytes by regulating the levels of TGF-β and pro-apoptotic proteins. Food and Chemical Toxicology.
Bovine Aortic Endothelial Cells: BAOEC
Nakamura, T., Yoshida, E., Fujie, T., Ogata, F., Yamamoto, C., Kawasaki, N. and Kaji, T., 2017. Synergistic cytotoxicity caused by forming a complex of copper and 2, 9-dimethyl-1, 10-phenanthroline in cultured vascular endothelial cells. The Journal of Toxicological Sciences, 42(6), pp.683-687.
Human Preadipocytes: HPAd
Zahid, H., Subbaramaiah, K., Iyengar, N.M., Zhou, X.K., Chen, I.C., Bhardwaj, P., Gucalp, A., Morrow, M., Hudis, C.A., Dannenberg, A.J. and Brown, K.A., 2017. Leptin regulation of the p53-HIF1α/PKM2-aromatase axis in breast adipose stromal cells—a novel mechanism for the obesity-breast cancer link. International Journal of Obesity. DOI: 10.1038/ijo.2017.273.
Human Pulmonary Artery Endothelial Cells: HPAEC
Nadeau, V., Potus, F., BOUCHERAT, O., Paradis, R., Tremblay, E., Iglarz, M., PAULIN, R., Bonnet, S. and PROVENCHER, S., 2017. Dual eta/etb blockade with macitentan improves both vascular remodelling and angiogenesis in pulmonary arterial hypertension. Pulmonary Circulation, p.2045893217741429.
Human Coronary Artery Endothelial Cells: HCAEC
Rai, R., Ghosh, A.K., Eren, M., Mackie, A.R., Levine, D.C., Kim, S.Y., Cedernaes, J., Ramirez, V., Procissi, D., Smith, L.H. and Woodruff, T.K., 2017. Downregulation of the Apelinergic Axis Accelerates Aging, whereas Its Systemic Restoration Improves the Mammalian Healthspan. Cell Reports, 21(6), pp.1471-1480.
MesoEndo Medium
Izadifar, M., Chapman, D., Babyn, P., Chen, X. and Kelly, M.E., 2017. UV-assisted 3D bioprinting of nano-reinforced hybrid cardiac patch for myocardial tissue engineering. Tissue Engineering, Part C Methods.
Human Cardiac Fibroblasts: HCF
Van Linthout, S., Hamdani, N., Miteva, K., Koschel, A., Müller, I., Pinzur, L., Aberman, Z., Pappritz, K., Linke, W.A. and Tschöpe, C., 2017. Placenta‐Derived Adherent Stromal Cells Improve Diabetes Mellitus‐Associated Left Ventricular Diastolic Performance. Stem cells translational medicine.
Duck, K.A., Simpson, I.A. and Connor, J.R., 2017. Regulatory mechanisms for iron transport across the blood-brain barrier. Biochemical and Biophysical Research Communications.
Human Preadipocyte Growth Medium
Zahid, H., Subbaramaiah, K., Iyengar, N.M., Zhou, X.K., Chen, I.C., Bhardwaj, P., Gucalp, A., Morrow, M., Hudis, C.A., Dannenberg, A.J. and Brown, K.A., 2017. Leptin regulation of the p53-HIF1α/PKM2-aromatase axis in breast adipose stromal cells—a novel mechanism for the obesity-breast cancer link. International Journal of Obesity. DOI: 10.1038/ijo.2017.273.
Human Peripheral Blood Mononuclear Cells: PBMC/HMNC-PB
Totani, T. and Tanaka, S., TOYO SEIKAN GROUP HOLDINGS, LTD., 2017. CULTURE CONTAINER AND METHOD FOR MANUFACTURING CULTURE CONTAINER. U.S. Patent 20,170,283,758.
Human Osteoblasts: Rheumatoid Arthritis: HOb-RA
Chen, Y.J., Chang, W.A., Hsu, Y.L., Chen, C.H. and Kuo, P.L., 2017. Deduction of Novel Genes Potentially Involved in Osteoblasts of Rheumatoid Arthritis Using Next-Generation Sequencing and Bioinformatic Approaches. International Journal of Molecular Sciences, 18(11), p.2396.
Anti-ERα 36 Ab
Yan, Y., Yu, L., Castro, L. and Dixon, D., 2017. ERα36, a variant of estrogen receptor α, is predominantly localized in mitochondria of human uterine smooth muscle and leiomyoma cells. PloS one, 12(10), p.e0186078.
Human Microvascular Endothelial Cell Media
Wu, Y., Zhang, Q. and Zhang, R., 2017. Kaempferol targets estrogen‑related receptor α and suppresses the angiogenesis of human retinal endothelial cells under high glucose conditions. Experimental and Therapeutic Medicine, 14(6), pp.5576-5582.
Human Lung Microvascular Endothelial Cells: HLMVEC
Iyer, R., Harris, J.F., Huang, J.H., Nath, P. and Przekwas, A., Los Alamos National Security, LLC, 2017. MULTI-ORGAN MEDIA COMPOSITIONS AND METHODS OF THEIR USE. U.S. Patent 20,170,275,587.
Classical Cell Media: MCDB-105
He, S., Niu, G., Shang, J., Deng, Y., Wan, Z., Zhang, C., You, Z. and Shen, H., 2017. The oncogenic Golgi phosphoprotein 3 like overexpression is associated with cisplatin resistance in ovarian carcinoma and activating the NF-κB signaling pathway. Journal of Experimental & Clinical Cancer Research, 36(1), p.137.
Human Umbilical Vein Endothelial Cells: HUVEC
Cao, X., Han, C., Wen, J., Guo, X. and Zhang, K., 2017. Nicotine increases apoptosis in HUVECs cultured in high glucose/high fat via Akt ubiquitination and degradation. Clinical and Experimental Pharmacology and Physiology.
Human Endothelial Cell Defined Medium
Rai, R., Ghosh, A.K., Eren, M., Mackie, A.R., Levine, D.C., Kim, S.Y., Cedernaes, J., Ramirez, V., Procissi, D., Smith, L.H. and Woodruff, T.K., 2017. Downregulation of the Apelinergic Axis Accelerates Aging, whereas Its Systemic Restoration Improves the Mammalian Healthspan. Cell Reports, 21(6), pp.1471-1480.
MesoEndo Medium
Zhou, T. and Chen, X., 2017. Long intergenic noncoding RNA p21 mediates oxidized LDL‑induced apoptosis and expression of LOX‑1 in human coronary artery endothelial cells. Molecular Medicine Reports, 16(6), pp.8513-8519.
Human Smooth Muscle Cell Media
Nadeau, V., Potus, F., BOUCHERAT, O., Paradis, R., Tremblay, E., Iglarz, M., PAULIN, R., Bonnet, S. and PROVENCHER, S., 2017. Dual eta/etb blockade with macitentan improves both vascular remodelling and angiogenesis in pulmonary arterial hypertension. Pulmonary Circulation, p.2045893217741429.
Anti-CD133
Choi, Y., Park, J., San Ko, Y., Kim, Y., Pyo, J.S., Jang, B.G., Kim, M.A., Lee, J.S., Chang, M.S. and Lee, B.L., 2017. FOXO1 reduces tumorsphere formation capacity and has crosstalk with LGR5 signaling in gastric cancer cells. Biochemical and Biophysical Research Communications, 493(3), pp.1349-1355.
Human Cardiac Fibroblast Basal Medium
Van Linthout, S., Hamdani, N., Miteva, K., Koschel, A., Müller, I., Pinzur, L., Aberman, Z., Pappritz, K., Linke, W.A. and Tschöpe, C., 2017. Placenta‐Derived Adherent Stromal Cells Improve Diabetes Mellitus‐Associated Left Ventricular Diastolic Performance. Stem cells translational medicine.
Human Umbilical Vein Endothelial Cells: HUVEC
Chen, X., Duong, M.N., Psaltis, P.J., Bursill, C.A. and Nicholls, S.J., 2017. High-density lipoproteins attenuate high glucose-impaired endothelial cell signaling and functions: potential implications for improved vascular repair in diabetes. Cardiovascular diabetology, 16(1), p.121.
Human Coronary Artery Endothelial Cells: HCAEC
Izadifar, M., Chapman, D., Babyn, P., Chen, X. and Kelly, M.E., 2017. UV-assisted 3D bioprinting of nano-reinforced hybrid cardiac patch for myocardial tissue engineering. Tissue Engineering, Part C Methods.
Rat Neural Stem Cell Differentiation Media
Hwang, M., Park, H.H., Choi, H., Lee, K.Y., Lee, Y.J. and Koh, S.H., 2017. Effects of aspirin and clopidogrel on neural stem cells. Cell Biology and Toxicology, pp.1-14.
Bovine Insulin
Zheng, Q., Bai, L., Zheng, S., Liu, M., Zhang, J., Wang, T., Xu, Z., Chen, Y., Li, J. and Duan, Z., 2017. Efficient inhibition of duck hepatitis B virus DNA by the CRISPR/Cas9 system. Molecular Medicine Reports, 16(5), pp.7199-7204.
Anti-ERα 36 Ab
Dai, Y.J., Qiu, Y.B., Jiang, R., Xu, M., Zhao, L., Chen, G.G. and Liu, Z.M., 2017. Concomitant high expression of ERα36, EGFR and HER2 is associated with aggressive behaviors of papillary thyroid carcinomas. Scientific Reports, 7(1), p.12279.
Human Umbilical Vein Endothelial Cells: HUVEC
Baimakhanov, Z., Sakai, Y., Yamanouchi, K., Hidaka, M., Soyama, A., Takatsuki, M. and Eguchi, S., 2017. Spontaneous hepatocyte migration towards an endothelial cell tube network. Journal of Tissue Engineering and Regenerative Medicine.
Rat Brain Microvascular Endothelial Cells: RBMVEC
Velasco-Aguirre, C., Morales-Zavala, F., Salas-Huenuleo, E., Gallardo-Toledo, E., Andonie, O., Muñoz, L., Rojas, X., Acosta, G., Sánchez-Navarro, M., Giralt, E. and Araya, E., 2017. Improving gold nanorod delivery to the central nervous system by conjugation to the shuttle Angiopep-2. Nanomedicine, 12(20), pp.2503-2517.
Attachment Factor Solution
Ruderisch, N., Schlatter, D., Kuglstatter, A., Guba, W., Huber, S., Cusulin, C., Benz, J., Rufer, A.C., Hoernschemeyer, J., Schweitzer, C. and Bülau, T., 2017. Potent and Selective BACE-1 Peptide Inhibitors Lower Brain Aβ Levels Mediated by Brain Shuttle Transport. EBioMedicine, 24, pp.76-92.
Human Smooth Muscle Cell Growth Medium
Yu, H., Jia, Q., Feng, X., Chen, H., Wang, L., Ni, X. and Kong, W., 2017. Hypoxia decrease expression of cartilage oligomeric matrix protein to promote phenotype switching of pulmonary arterial smooth muscle cells. The International Journal of Biochemistry & Cell Biology, 91, pp.37-44.
Anti-CD133
Cho, Y.C., Nguyen, T.T., Park, S.Y., Kim, K., Kim, H.S., Jeong, H.G., Kim, K.K. and Kim, H., 2017. Bromopropane Compounds Increase the Stemness of Colorectal Cancer Cells. International Journal of Molecular Sciences, 18(9), p.1888.
Human Coronary Artery Endothelial Cells: HCAEC
Zhou, T. and Chen, X., 2017. Long intergenic noncoding RNA p21 mediates oxidized LDL‑induced apoptosis and expression of LOX‑1 in human coronary artery endothelial cells. Molecular Medicine Reports, 16(6), pp.8513-8519.
Human Umbilical Vein Endothelial Cells: HUVEC
Lai, C.J., Cheng, H.C., Lin, C.Y., Huang, S.H., Chen, T.H., Chung, C.J., Chang, C.H., Wang, H.D. and Chuu, C.P., 2017. Activation of liver X receptor suppresses angiogenesis via induction of ApoD. The FASEB Journal, pp.fj-201700374R.
Human Osteoblasts: HOb
Chen, Y.J., Chang, W.A., Hsu, Y.L., Chen, C.H. and Kuo, P.L., 2017. Deduction of Novel Genes Potentially Involved in Osteoblasts of Rheumatoid Arthritis Using Next-Generation Sequencing and Bioinformatic Approaches. International Journal of Molecular Sciences, 18(11), p.2396.
Human Aortic Endothelial Cells: HAOEC
Lo, W. Y., Peng, C. T., & Wang, H. J. (2017). MicroRNA-146a-5p Mediates High Glucose-Induced Endothelial Inflammation via Targeting Interleukin-1 Receptor-Associated Kinase 1 Expression. Frontiers in Physiology, 8, 551.
Human Chondrocytes: HC
Bellayr, I.H., Kumar, A. and Puri, R.K., 2017. MicroRNA expression in bone marrow-derived human multipotent Stromal cells. BMC Genomics, 18(1), p.605.
Rat aortic smooth muscle cells (RASMC)
Chuang, T.D. and Khorram, O., 2017. Glucocorticoids regulate MiR-29c levels in vascular smooth muscle cells through transcriptional and epigenetic mechanisms. Life Sciences, 186, pp.87-91.
Human Pulmonary Artery Smooth Muscle Cells: HPASMC
Chakraborti, S., Sarkar, J., Bhuyan, R. and Chakraborti, T., 2017. Role of curcumin in PLD activation by Arf6-cytohesin1 signaling axis in U46619-stimulated pulmonary artery smooth muscle cells. Molecular and Cellular Biochemistry, pp.1-13.
Human Mesenchymal Stem Cells: HMSC
Janda, C.Y., Dang, L.T., You, C., Chang, J., De Lau, W., Zhong, Z.A., Yan, K.S., Marecic, O., Siepe, D., Li, X. and Moody, J.D et al. 2017. Surrogate Wnt agonists that phenocopy canonical Wnt and β-catenin signalling. Nature, 545(7653), pp.234-237.
Human Aortic Smooth Muscle Cells: HAOSMC
Jiang, W., Wang, Z., Hu, Z., Wu, H., Hu, R., Hu, X., Ren, Z. and Huang, J., 2017. Blocking the ERK1/2 signal pathway can inhibit S100A12 induced human aortic smooth muscle cells damage. Cell Biology International. DOI: 10.1002/cbin.10840
Human Pulmonary Artery Smooth Muscle Cells: HPASMC
Chakraborti, S., Sarkar, J., Chowdhury, A. and Chakraborti, T., 2017. Role of ADP ribosylation factor6− Cytohesin1− PhospholipaseD signaling axis in U46619 induced activation of NADPH oxidase in pulmonary artery smooth muscle cell membrane. Archives of Biochemistry and Biophysics. DOI: 10.1016/j.abb.2017.08.012
Human Dermal Fibroblasts: HDF
Bellayr, I.H., Kumar, A. and Puri, R.K., 2017. MicroRNA expression in bone marrow-derived human multipotent Stromal cells. BMC Genomics, 18(1), p.605.
Human Endothelial Cell Growth Medium
Lo, W. Y., Peng, C. T., & Wang, H. J. (2017). MicroRNA-146a-5p Mediates High Glucose-Induced Endothelial Inflammation via Targeting Interleukin-1 Receptor-Associated Kinase 1 Expression. Frontiers in Physiology, 8, 551.
Rat Dermal Fibroblasts: RDF
Uchinaka A, Kawaguchi N, Ban T, Hamada Y, Mori S, Maeno Y, Sawa Y, Nagata K, Yamamoto H. 2017. Evaluation of dermal wound healing activity of synthetic peptide SVVYGLR. Biochem Biophys Res Commun. 2017 Jul 24. pii: S0006-291X(17)31482-1.
Endothelial Cell Growth Media Kit
Y Xue, CS Hilaire, L Hortells, JA Phillippi, V Sant, S Sant. 2017. Shape-Specific Nanoceria Mitigate Oxidative Stress-Induced Calcification in Primary Human Valvular Interstitial Cell Culture. Cellular and Molecular Bioengineering, 1-18.
Human Mesenchymal Stem Cells: HMSC
Bellayr, I.H., Kumar, A. and Puri, R.K., 2017. MicroRNA expression in bone marrow-derived human multipotent Stromal cells. BMC Genomics, 18(1), p.605.
MesoEndo Medium
Izadifar M, Babyn P, Kelly ME, Chapman D, Chen X. 2017. Bioprinting pattern-dependent electrical/mechanical behavior of cardiac alginate implants: characterization and ex-vivo phase-contrast microtomography assessment. Tissue Eng Part C Methods. doi: 10.1089/ten.TEC.2017.0222.
Osteogenic and Adipogenic Canine Differentiation Media
Matsuda, T., Takami, T., Sasaki, R., Nishimura, T., Aibe, Y., Paredes, B. D., Quintanilha, L. F., Matsumoto, T., Ishikawa, T., Yamamoto, N., Tani, K., Terai, S., Taura, Y. and Sakaida, I. 2017. A canine liver fibrosis model to develop a therapy for liver cirrhosis using cultured bone marrow-derived cells. Hepatology Communications. doi:10.1002/hep4.1071.
Human Coronary Artery Endothelial Cells: HCAEC
Izadifar M, Babyn P, Kelly ME, Chapman D, Chen X. 2017. Bioprinting pattern-dependent electrical/mechanical behavior of cardiac alginate implants: characterization and ex-vivo phase-contrast microtomography assessment. Tissue Eng Part C Methods. doi: 10.1089/ten.TEC.2017.0222.
Bovine Insulin
Luchun Li, Yan Li, Lulu Wang, Zhijuan Wu, Huiwen Ma, Jianghe Shao, Dairong Li, Huiqing Yu, Weiqi Nian, Donglin Wang. 2017. Inhibition of Hes1 enhances lapatinib sensitivity in gastric cancer sphere-forming cells. Oncology Letters. https://doi.org/10.3892/ol.2017.6683.
Human Osteoblasts: Hob
Bellayr, I.H., Kumar, A. and Puri, R.K., 2017. MicroRNA expression in bone marrow-derived human multipotent Stromal cells. BMC Genomics, 18(1), p.605.
Rat Hippocampal Neurons: RHiN
McDonough Patrick M., Prigozhina Natalie L., Basa Ranor C.B., and Price Jeffrey H. 2017. Assay of Calcium Transients and Synapses in Rat Hippocampal Neurons by Kinetic Image Cytometry and High-Content Analysis: An In Vitro Model System for Postchemotherapy Cognitive Impairment. ASSAY and Drug Development Technologies. 15(5): 220-236.
Human Dermal Fibroblasts: HDF
Y Esparza, A Ullah, Y Boluk, J Wu. 2017. Preparation and characterization of thermally crosslinked poly (vinyl alcohol)/feather keratin nanofiber scaffolds. Materials & Design, https://doi.org/10.1016/j.matdes.2017.07.052.
Canine Osteoblasts: CnOb
Troyer RM, Ruby CE, Goodall CP, Yang L, Maier CS, Albarqi HA, Brady JV, Bathke K, Taratula O, Mourich D, Bracha S. 2017. Exosomes from Osteosarcoma and normal osteoblast differ in proteomic cargo and immunomodulatory effects on T cells. Exp Cell Res. pii: S0014-4827(17)30365-8.
Human Carotid Artery Endothelial Cells: HCtAEC
Pott, G. B., Tsurudome, M., Bui, J., Banfield, C., Hourieh, S., Pratap, H., & Goalstone, M. L. 2017. VCAM-1 Mediates Cigarette Smoke Extract Enhancement of Monocyte Adhesion to Human Carotid Endothelial Cells. Medical Research Archives. Volume 5, issue 7.
Human Umbilical Vein Endothelial Cells: HUVEC
Goszcz, K., Deakin, S., Duthie, G. G., Stewart, D., Megson, I. L., & Megson, I. L. 2017. Bioavailable concentrations of delphinidin and its metabolite, gallic acid, induce antioxidant protection associated with increased intracellular glutathione in cultured endothelial cells. Oxidative Medicine and Cellular Longevity.
Rat Endothelial Cell Basal Medium
Iba, T., Sasaki, T., Ohshima, K., Sato, K., Nagaoka, I., Thachil, J., Bucur, S.Z., Levy, J.H., Despotis, G.J., Spiess, B.D. and Hillyer, C.D., 2017. The comparison of the protective effects of α-and β-antithrombin against vascular endothelial cell damage induced by histone in vitro. TH Open, 1(01), pp.e3-e10.
Human Dermal Fibroblasts: HDF
Martinez-Cerdeno, Veronica, Bonnie Barrilleaux, Ashley McDonough, Jeanelle Ariza, Benjamin Yuen, Priyanka Somanath, Catherine Le, Craig Steward, Kayla Horton, and Paul Knoepfler. 2017. Behavior of xeno-transplanted undifferentiated human induced pluripotent stem cells is impacted by microenvironment without evidence of tumors. Stem Cells and Development. https://doi.org/10.1089/scd.2017.0059.
Anti-CD133
Phiboonchaiyanan, P.P. and Chanvorachote, P., 2017. Suppression of a cancer stem-like phenotype mediated by alpha-lipoic acid in human lung cancer cells through down-regulation of β-catenin and Oct-4. Cellular Oncology, pp.1-14.
Bovine Aortic Endothelial Cells: BAOEC
Dang, L.T., Aburatani, T., Marsh, G.A., Johnson, B.G., Alimperti, S., Yoon, C.J., Huang, A., Szak, S., Nakagawa, N., Gomez, I. and Ren, S., 2017. Hyperactive FOXO1 results in lack of tip stalk identity and deficient microvascular regeneration during kidney injury. Biomaterials. https://doi.org/10.1016/j.biomaterials.2017.07.010
Rat Aortic Endothelial Cells: RAOEC
Iba, T., Sasaki, T., Ohshima, K., Sato, K., Nagaoka, I., Thachil, J., Bucur, S.Z., Levy, J.H., Despotis, G.J., Spiess, B.D. and Hillyer, C.D., 2017. The comparison of the protective effects of α-and β-antithrombin against vascular endothelial cell damage induced by histone in vitro. TH Open, 1(01), pp.e3-e10.
Human Smooth Muscle Cell Media
Vanags, L.Z., Tan, J.T., Santos, M., Michael, P.S., Ali, Z., Bilek, M.M., Wise, S.G. and Bursill, C.A., 2017. Plasma activated coating immobilizes apolipoprotein AI to stainless steel surfaces in its bioactive form and enhances biocompatibility. Nanomedicine: Nanotechnology, Biology and Medicine. https://doi.org/10.1016/j.nano.2017.06.012.
Bovine Aortic Endothelial Cells: BAOEC
Berger, A.J., Linsmeier, K., Kreeger, P.K. and Masters, K.S., 2017. Decoupling the effects of stiffness and fiber density on cellular behaviors via an interpenetrating network of gelatin-methacrylate and collagen. Biomaterials. https://doi.org/10.1016/j.biomaterials.
Human Adipocyte Differentiation Medium
Dong, Y., Betancourt, A., Belfort, M. and Yallampalli, C., 2017. Targeting Adrenomedullin to Improve Lipid Homeostasis in Diabetic Pregnancies. The Journal of Clinical Endocrinology & Metabolism. https://doi.org/10.1210/jc.2017-00920.
Human Umbilical Vein Smooth Muscle Cells: HUVSMC
Vanags, L.Z., Tan, J.T., Santos, M., Michael, P.S., Ali, Z., Bilek, M.M., Wise, S.G. and Bursill, C.A., 2017. Plasma activated coating immobilizes apolipoprotein AI to stainless steel surfaces in its bioactive form and enhances biocompatibility. Nanomedicine: Nanotechnology, Biology and Medicine. https://doi.org/10.1016/j.nano.2017.06.012.
Rat Brain Microvascular Endothelial Cells: RBMVEC
Gray, S.M., Aylor, K.W. and Barrett, E.J., 2017. Unravelling the regulation of insulin transport across the brain endothelial cell. Diabetologia, pp.1-10.
HOb medium
Canal, C., Fontelo, R., Hamouda, I., Guillem-Marti, J., Cvelbar, U. and Ginebra, M.P., 2017. Plasma-induced selectivity in bone cancer cells death. Free Radical Biology and Medicine. https://doi.org/10.1016/j.freeradbiomed.2017.05.023.
Human Dermal Microvascular Endothelial Cells: CADMEC/HMVEC
Tan, W., Wang, J., Zhou, F., Gao, L., Rong, Y., Liu, H., Sukanthanag, A., Wang, G., Mihm, M.C., Chen, D.B. and Nelson, J.S., 2017. Coexistence of EphB1 and EphrinB2 in Port Wine Stain Endothelial Progenitor Cells Contributes to Clinicopathological Vasculature Dilatation. British Journal of Dermatology. DOI: 10.1111/bjd.15716.
Human Mesenchymal Stem Cell Media
Bellayr, I.H., Kumar, A. and Puri, R.K., 2017. MicroRNA expression in bone marrow-derived human multipotent Stromal cells. BMC Genomics, 18(1), p.605.
Human Endothelial Cell Media
Tan, W., Wang, J., Zhou, F., Gao, L., Rong, Y., Liu, H., Sukanthanag, A., Wang, G., Mihm, M.C., Chen, D.B. and Nelson, J.S., 2017. Coexistence of EphB1 and EphrinB2 in Port Wine Stain Endothelial Progenitor Cells Contributes to Clinicopathological Vasculature Dilatation. British Journal of Dermatology. DOI: 10.1111/bjd.15716.
Human Chondrocytes: Osteoarthritis: HC-OA
Rosenberg, J.H., Rai, V., Dilisio, M.F., Sekundiak, T.D. and Agrawal, D.K., 2017. Increased expression of damage-associated molecular patterns (DAMPs) in osteoarthritis of human knee joint compared to hip joint. Molecular and Cellular Biochemistry, pp.1-11.
Human Endothelial Cell Media
Shatanawi, A. and Momani, M.S., 2017. Plasma arginase activity is elevated in type 2 diabetic patients. Biomedical Research, 28(9).
Smooth Muscle Cells
Kikuchi, S., Chen, L., Xiong, K., Saito, Y., Azuma, N., Tang, G., Sobel, M., Wight, T.N. and Kenagy, R.D., 2017. Smooth muscle cells of human veins show an increased response to injury at valve sites. Journal of Vascular Surgery. http://dx.doi.org/10.1016/j.jvs.2017.03.447.
Human Osteoblasts: HOb
Canal, C., Fontelo, R., Hamouda, I., Guillem-Marti, J., Cvelbar, U. and Ginebra, M.P., 2017. Plasma-induced selectivity in bone cancer cells death. Free Radical Biology and Medicine. https://doi.org/10.1016/j.freeradbiomed.2017.05.023.
Human Fibroblast-Like Synoviocytes: Rheumatoid Arthritis: HFLS-RA
Wang, S., Liang, S., Zhao, X., He, Y. and Qi, Y., 2017. Propofol inhibits cell proliferation and invasion in rheumatoid arthritis fibroblast-like synoviocytes via the nuclear factor-κB pathway. American journal of translational research, 9(5), p.2429.
Bovine Aortic Endothelial Cells: BAOEC
Shatanawi, A. and Momani, M.S., 2017. Plasma arginase activity is elevated in type 2 diabetic patients. Biomedical Research, 28(9).
Dou, P., R. Hu, W. Zhu, Q. Tang, D Li, H. Li and W. Wang. 2017. Long non-coding RNA HOTAIR promotes expression of ADAMTS-5 in human osteoarthritic articular chondrocytes. Die Pharmazie, 72:113-117.
Dou, P., R. Hu, W. Zhu, Q. Tang, D Li, H. Li and W. Wang. 2017. Long non-coding RNA HOTAIR promotes expression of ADAMTS-5 in human osteoarthritic articular chondrocytes. Die Pharmazie, 72:113-117.
Dou, P., R. Hu, W. Zhu, Q. Tang, D Li, H. Li and W. Wang. 2017.Long non-coding RNA HOTAIR promotes expression of ADAMTS-5 in human osteoarthritic articular chondrocytes. Die Pharmazie, 72:113-117.
Zhao, G., X. Cheng, L. Piao, L. Hu, Y. Lei, G. Yang, A. Inoue, S. Ogasawara, H. Wu, N. Hao, K. Okumara and M. Kuzuya. 2017. The Soluble VEGF Receptor sFlt-1 Contributes to Impaired Neovascularization in Aged Mice. Aging and Disease, 8(3).
FDA, US Food and Drug Administration; EMA, European Medicines Agency; HEK, human embryonic kidney; NA, not approved; rFVIIIFc, recombinant factor VIII Fc fusion protein; rFIXFc, recombinant factor IX Fc fusion protein; rhFVIII, recombinant human factor VIII.
aData obtained from publically available resources (October 2014); all approved products may not be included.
bReferences: (ALPROLIX®, 2014; Bakker et al., 2005; Behrens et al., 2014; Casademunt et al., 2012; DYNEPO®, 2007; ELAPRASE®, 2012, 2013; ELOCTATE®, 2014; European Medicines Agency and Committee for Medicinal Products for Human Use, 2014; Glaesner et al., 2010; Octapharma, 2014; REPLAGAL®, 2006; TRULICITY™, 2014; VPRIV®, 2010a,b; XIGRIS®, 2008).
Table 3.
Comparison of human cell lines with other expression systems in the production of therapeutic proteins.
Biotherapeutic proteins represent a mainstay of treatment for a multitude of conditions, for example, autoimmune disorders, hematologic disorders, hormonal dysregulation, cancers, infectious diseases and genetic disorders. The technologies behind their production have changed substantially since biotherapeutic proteins were first approved in the 1980s. Although most biotherapeutic proteins developed to date have been produced using the mammalian Chinese hamster ovary and murine myeloma (NS0, Sp2/0) cell lines, there has been a recent shift toward the use of human cell lines. One of the most important advantages of using human cell lines for protein production is the greater likelihood that the resulting recombinant protein will bear post-translational modifications (PTMs) that are consistent with those seen on endogenous human proteins. Although other mammalian cell lines can produce PTMs similar to human cells, they also produce non-human PTMs, such as galactose-α1,3-galactose and N-glycolylneuraminic acid, which are potentially immunogenic. In addition, human cell lines are grown easily in a serum-free suspension culture, reproduce rapidly and have efficient protein production. A possible disadvantage of using human cell lines is the potential for human-specific viral contamination, although this risk can be mitigated with multiple viral inactivation or clearance steps. In addition, while human cell lines are currently widely used for biopharmaceutical research, vaccine production and production of some licensed protein therapeutics, there is a relative paucity of clinical experience with human cell lines because they have only recently begun to be used for the manufacture of proteins (compared with other types of cell lines). With additional research investment, human cell lines may be further optimized for routine commercial production of a broader range of biotherapeutic proteins.
Protein therapeutics (including monoclonal antibodies [mAbs], peptides and recombinant proteins) represent the largest group of new products in development by the biopharmaceutical industry (Durocher & Butler, 2009; Ho & Chien, 2014).
These products are produced in a wide variety of platforms, including non-mammalian expression systems (bacterial, yeast, plant and insect) and mammalian expression systems (including human cell lines) (Ghaderi et al., 2012). Importantly, the most appropriate expression system depends on the particular protein to be expressed. Mammalian expression systems are generally the preferred platform for manufacturing biopharmaceuticals, as these cell lines are able to produce large, complex proteins with post-translational modifications (PTMs; most notably glycosylation) similar to those produced in humans (Durocher & Butler, 2009; Ghaderi et al., 2012; Swiech et al., 2012). Moreover, in the case of mammalian cell lines, and animal cell lines in general, most proteins can be secreted rather than requiring cell lysis to extract with subsequent protein refolding (as is the case with bacteria/prokaryotes). The most common mammalian (non-human) cell lines used for therapeutic protein production include Chinese hamster ovary (CHO) cells, baby hamster kidney (BHK21) cells and murine myeloma cells (NS0 and Sp2/0) (Estes & Melville, 2014). However, these non-human mammalian cell lines also produce PTMs that are not expressed in humans, namely galactose-α1,3-galactose (α-gal) and N-glycolylneuraminic acid (NGNA). Because humans possess circulating antibodies against both of these N-glycans, non-human cell lines are usually screened during their production to identify clones with acceptable glycan profiles (Ghaderi et al., 2010).
Human cell lines have the ability to produce proteins most similar to those synthesized naturally in humans, which may be an advantage compared with other mammalian expression systems (Ghaderi et al., 2010). In particular, the structure, number and location of post-translational N-glycans can affect the biologic activity, protein stability, clearance rate and immunogenicity of biotherapeutic proteins (Arnold et al., 2007; Ghaderi et al., 2010; Swiech et al., 2012).
The first human cell line, HeLa, was established in 1951 from a cervical cancer (Scherer et al., 1953). Human diploid cells were developed in the 1960s for vaccine manufacturing; however, concerns for a latent oncogenic agent in these cell lines (despite a lack of suggestive phenotypic characteristics) delayed their acceptance. Currently, human diploid cells are used in the manufacture of many viral vaccines (Petricciani & Sheets, 2008). However, due to their rapid growth, high protein yield, and the investment in system optimization, animal cells remained the substrate of choice for the production of recombinant proteins and mAbs (Petricciani & Sheets, 2008).
Today, advances in technology have allowed for increased productivity with human cell lines, and there are now approved recombinant biotherapeutic products produced from the human embryonic kidney 293 (HEK293) and fibrosarcoma HT-1080 cell lines (Beck, 2009; Casademunt et al., 2012; Dumont et al., 2012; Glaesner et al., 2010; Peters et al., 2010; Wraith, 2008; Zimran et al., 2013). Additional biotherapeutic products produced in the PER.C6, HKB-11, CAP and HuH-7 human cell lines are currently being evaluated (Enjolras et al., 2012; Estes & Melville, 2014; Jones et al., 2003; Mei et al., 2006; Swiech et al., 2011, 2015). This article is a narrative review of the cell lines (with a focus on human cell lines) used for production of biotherapeutic proteins, both approved and in development.
Non-human expression systems used to manufacture biotherapeutic products
Many non-human expression systems have been utilized in the production of currently approved biotherapeutic proteins (Table 1).
Table 1.
Non-human expression systems used in the production of biotherapeutics approved in the United States and Europea,b.
Bacterial expression systems (e.g. Escherichia coli) possess the advantages of being straightforward to culture, with rapid cell growth and high yields. In addition, protein expression can be initiated through promoter induction by addition of lactose or the lactose analogue isopropyl-β-d-thiogalactopyranoside (IPTG; IPTG induces the promoters lac, tac and trc). However, such systems are unable to produce complex, mammalian-like glycosylation due to the absence of the necessary enzymatic components and the intracellular compartmentalization required (Ghaderi et al., 2012; Graumann & Premstaller, 2006). In addition, mammalian proteins produced in these systems often aggregate, forming inclusion bodies, due to the low solubility of mammalian proteins in prokaryotic cells and absence of appropriate protein chaperone systems. Proteins produced in bacterial expression systems must often be extracted from inclusion bodies and refolded. Bacterial systems are therefore generally used for production of non-glycosylated proteins, including some mAbs, hormones, cytokines and enzymes (Ghaderi et al., 2012; Graumann & Premstaller, 2006).
Similar to bacterial expression systems, yeast expression systems (e.g. Saccharomyces cerevisiae and Pichia pastoris) achieve rapid cell growth and high-protein yields with straightforward production scalability and without the need for animal-derived growth factors (Gerngross, 2004). Yeast cell lines may also be used to produce proteins that cannot be obtained from E. coli due to the problems associated with folding and stereochemistry (Gerngross, 2004). The key challenge associated with yeast expression systems is their production of high mannose residues within their expressed PTMs (50–200 vs three molecules in human cells, as part of either N– or O-linked glycan structures), which may confer a short half-life and render proteins less efficacious and even immunogenic in humans (Dean, 1999; Gemmill & Trimble, 1999; Gerngross, 2004; Lam et al., 2007; Mochizuki et al., 2001). The development of yeasts that have been genetically modified to address the issue of high mannose content has been reported (Chiba et al., 1998; Gerngross, 2004; Ghaderi et al., 2012; Hamilton et al., 2003). The expression of a fully humanized sialylated glycoprotein in glycoengineered yeast constitutes a major advance in the use of yeast expression systems for biopharmaceutical manufacturing (Hamilton & Gerngross, 2007).
Plant and insect cell expression systems are able to produce proteins with complex glycosylation patterns; however, the glycan structures produced are significantly different from those produced in humans (Ghaderi et al., 2012). Plants lack many of the key glycosylated residues present in humans, most notably sialic acids. In addition, they produce α1,3-fructose and β1,2-xylose, which are absent in humans and may be immunogenic (Ghaderi et al., 2012). Notably, in 2012, taliglucerase alfa (ELELYSO®; Pfizer, New York, NY) was approved by the US Food and Drug Administration (FDA) for the treatment of type 1 Gaucher disease. This therapy is produced using genetically modified carrot plant root cells that produce the enzyme with a human compatible glycan profile (ELELYSO™, 2014).
Insect cells infected with the viral vector baculovirus (baculovirus-insect cell expression system) can also efficiently express recombinant proteins, and these systems are mostly used for the development of virus-like particles and, subsequently, vaccines (Kost et al., 2005; Liu et al., 2013). However, although they produce N-glycan precursors, these are trimmed, resulting in either high mannose or paucimannose residues that do not develop further into terminal galactose and/or sialic acid residues (Kost et al., 2005). This is evidenced by the lack of either galactosyltransferase or sialyltransferase activity. As in plants, insect systems may also express the fucosylated α1,3-linkage (Staudacher et al., 1999). However, in recent years, there have been developments in the use of transgenic insect cells, with humanized protein glycosylation mechanisms (Kost et al., 2005).
The majority of currently licensed biotherapeutic products are produced in non-human mammalian expression systems (Table 1), as these systems are able to produce PTMs that (outside of a human expression system) most closely resemble those in humans (Ghaderi et al., 2010). These expression systems are used to produce mAbs, hormones, cytokines, enzymes and clotting factors (Ghaderi et al., 2012).
The most frequently used mammalian system is the CHO cell line, which is used in the manufacture of >70% of currently approved recombinant proteins (Butler & Spearman, 2014). This cell line has demonstrated several major advantages. First, CHO cells are able to grow in suspension culture (which enables large-scale production; other cell lines, such as insect cells, also have this ability) and serum-free chemically defined media (enabling reproducibility across different batches of cultures with a better safety profile than in media that contain human- or animal-derived proteins) (Kim et al., 2012; Lai et al., 2013; Rossi et al., 2012). Historically, CHO cells allowed gene amplification, resulting in a higher recombinant protein yield (up to the gram per liter range for some proteins) and specific productivity, which was previously an issue in other mammalian cell lines (Carlage et al., 2012; Kim et al., 2012; Yang et al., 2014a,b). Other advances, such as the creation of stronger expression units and advanced hosts, better selection strategies (e.g. through technologic advances in screening for high-productivity clones) and targeting the transgene to transcriptional hotspots (site-specific integration of transgenes), also contribute to the high protein yields attained from these cells (Kim et al., 2012). In addition, this expression system is highly tolerant to changes in pH, oxygen level, pressure or temperature during manufacturing (Ghaderi et al., 2012; Lai et al., 2013). Furthermore, due to the long period of time that this cell line has been used, there is a degree of familiarity with the CHO platform within development and manufacturing organizations, regulatory agencies, and suppliers (e.g. cell culture media suppliers), which could potentially decrease overall timelines. This familiarity may also be beneficial when assessing contaminant profiles (e.g. host cell proteins), which may be better characterized for CHO cells compared with newer cell lines.
The first recombinant biotherapeutic protein produced in CHO cells was tissue plasminogen activator, approved in 1986 (Kim et al., 2012). Therefore, the safety profile of CHO cells has been established for more than 20 years (Butler & Spearman, 2014; Kim et al., 2012). CHO cells have been shown to have reduced susceptibility to certain viral infections compared with other mammalian cell lines (e.g. the BHK cell line), and routine screening systems for adventitious agents are effective in detecting cell line infections (Berting et al., 2010). This reduced susceptibility may be due to the fact that many viral entry genes are not expressed in CHO cells (Xu et al., 2011). Further, there is perceived species barrier protection with the use of hamster-derived cells, reducing the potential risk of transfer of contaminating adventitious agents to humans (Berting et al., 2010; Swiech et al., 2012). However, many viruses have the ability to cross the species barrier and may still pose a risk (Pauwels et al., 2007).
Perhaps the most important advantage of CHO cells is that they are able to produce proteins with complex bioactive PTMs that are similar to those produced in humans. However, CHO cells are unable to produce some types of human glycosylation (CHO cells lack α[2-6] sialyltransferase α[1-3/4] fucosyltransferases) and they produce glycans that are not expressed in humans, namely α-gal and NGNA (Bosques et al., 2010; Dietmair et al., 2012; Ghaderi et al., 2012). Circulating antibodies against both of these N-glycans are present in humans, which may lead to increased immunogenicity and altered pharmacokinetics of these products when used in humans (Ghaderi et al., 2010; Padler-Karavani et al., 2008). Additional screening in CHO cells is required in order to isolate clones lacking the α-gal and NGNA glycans. This screening may result in otherwise productive clones needing to be discarded (Ghaderi et al., 2010). However, the attachment of non-human glycans may not be a concern for therapeutic proteins that do not require glycosylation, which illustrates the importance of considering the specific product molecule when choosing an appropriate cell line for production of a protein.
Other mammalian cell lines used for the production of biotherapeutic proteins include BHK-21 cells, used in the production of some coagulation factors such as factor VIII (Wurm, 2004). When murine myeloma cell lines (NS0 and Sp2/0) have been used historically, they have generally been used in the production of mAbs, for example, palivizumab and ofatumumab (Barnes et al., 2000; Butler & Spearman, 2014; Ghaderi et al., 2012). These myeloma cells were derived from immunoglobulin-producing tumor cells that no longer produced their original immunoglobulins; these cells possess the appropriate machinery for producing and secreting these proteins (Barnes et al., 2000).
For proteins produced in all of these non-human cell lines, as well as those produced in human cell lines, potential safety concerns arise from the possibility of process-related contaminants and immunogenicity (World Health Organization, 2013). Process-related contaminants may include infectious agents (viral, bacterial, fungal, etc.) with the potential to result in host infection, nucleic acid contaminants with the potential to integrate into the host genome (theoretical), and other contaminants from the manufacturing process, such as exogenous non-human epitopes (e.g. from animal serum used during the manufacturing process) that can be incorporated into human cells and the resultant biotherapeutic protein (Ghaderi et al., 2012).
Human cell lines used to manufacture licensed products
HEK293 and HT-1080 are the two human cell lines most often used in the production of biotherapeutic proteins, which offer the advantage of producing fully human PTMs (Tables 2 and and3)3) (Loignon et al., 2008; Swiech et al., 2012).
Table 2.
Human cells lines and their therapeutic protein productsa,b.
FDA, US Food and Drug Administration; EMA, European Medicines Agency; HEK, human embryonic kidney; NA, not approved; rFVIIIFc, recombinant factor VIII Fc fusion protein; rFIXFc, recombinant factor IX Fc fusion protein; rhFVIII, recombinant human factor VIII.
aData obtained from publically available resources (October 2014); all approved products may not be included.
bReferences: (ALPROLIX®, 2014; Bakker et al., 2005; Behrens et al., 2014; Casademunt et al., 2012; DYNEPO®, 2007; ELAPRASE®, 2012, 2013; ELOCTATE®, 2014; European Medicines Agency and Committee for Medicinal Products for Human Use, 2014; Glaesner et al., 2010; Octapharma, 2014; REPLAGAL®, 2006; TRULICITY™, 2014; VPRIV®, 2010a,b; XIGRIS®, 2008).
Table 3.
Comparison of human cell lines with other expression systems in the production of therapeutic proteins.
Advantages
Disadvantages
• Absence of potentially immunogenic PTMs due to human-compatible glycosylation • Easily grown in suspension serum-free culture • Achieve rapid reproduction • Amenable to a number of transfection methods
• Clinical experience is not as extensive as for other cell lines, although experience is growing • Potential susceptibility to human viral contamination
HEK293 cells are easily grown in suspension serum-free culture, reproduce rapidly, are amenable to a number of transfection methods, and are highly efficient at protein production (Swiech et al., 2012; Thomas & Smart, 2005).
HEK293-H (Berkner, 1993) and 293-F (Vink et al., 2014) cell lines are clonal isolates of the HEK293 cell line that were selected for fast growth in serum-free medium, superior transfection efficiency, and a high level of protein production (Gibco, 2014). Subclone 293-H also has improved adherence to monolayer culture (when serum-supplemented media are used) compared with other cell lines. Other modified HEK293 cells include the HEK293-T cell line and HEK293-EBNA1 cells. The HEK293-T (293-T) cell line expresses the simian virus 40 large T antigen and is capable of expressing high titers of viral gene vectors for use in gene therapy (Yamaguchi et al., 2003). HEK293-T cells are often used for the production of retroviral vectors (Yamaguchi et al., 2003). HEK293-EBNA1 cells stably express the Epstein-Barr virus EBNA-1 gene, controlled by the cytomegalovirus promoter and demonstrate a greater growth rate and maximal cell density relative to parental HEK293 cells (Schlaeger & Christensen, 1999).
HEK293 cells have been widely used to produce research-grade proteins for many years and, more recently, five therapeutic agents produced in HEK293 cells have been approved by the FDA or the European Medicines Agency (EMA) for therapeutic use. These agents are drotrecogin alfa (XIGRIS®; Eli Lilly Corporation, Indianapolis, IN), recombinant factor IX Fc fusion protein (rFIXFc; Biogen, Cambridge, MA), recombinant factor VIII Fc fusion protein (rFVIIIFc; Biogen, Cambridge, MA), human cell line recombinant factor VIII (human-cl rhFVIII; NUWIQ®; Octapharma, Lachen, Switzerland) and dulaglutide (TRULICITY®; Eli Lily, Indianapolis, IN).
Drotrecogin alfa is a recombinant activated protein C that was approved by the FDA in 2001 and by the EMA in 2002 for the treatment of patients with severe sepsis. HEK293 cells were chosen by the manufacturer for production of drotrecogin alfa because its activity required two PTMs, propeptide cleavage and γ-carboxylation of its glutamic acid residues, which CHO cells cannot produce with adequate efficiency (Berkner, 1993; Durocher & Butler, 2009). The product was approved (Bernard et al., 2001), but was later voluntarily withdrawn from the market by its manufacturer (Eli Lilly) in 2011 following the randomized placebo-controlled Prospective Recombinant Human Activated Protein C Worldwide Evaluation in Severe Sepsis and Septic Shock (PROWESS-SHOCK) trial, which demonstrated no mortality benefit with drotrecogin alfa compared with placebo for patients experiencing septic shock (Green et al., 2012; Ranieri et al., 2012).
rFVIIIFc and rFIXFc are recombinant fusion proteins that were approved by the FDA in 2014 for the control and prevention of bleeding episodes, perioperative management and routine prophylaxis to prevent or reduce the frequency of bleeding episodes in people with hemophilia A and B, respectively (ALPROLIX®, 2014; ELOCTATE®, 2014; Mahlangu et al., 2014; Powell et al., 2013). They are also approved in Canada, Australia and Japan. rFVIIIFc consists of B domain–deleted recombinant factor VIII genetically fused to the Fc portion of immunoglobulin G1 (IgG1) and is produced in HEK293-H cells (Dumont et al., 2012; ELOCTATE®, 2014; Peters et al., 2013). The rFVIIIFc fed-batch culture process is robust at scales of 200, 2000 and 15 000 liters, with the potential for a second-generation process to achieve even higher cell densities, on the order of 3.5 × 107 vc/ml (Huang et al., 2014). rFIXFc was also produced using HEK293-H cells, and consists of the factor IX sequence covalently linked to the Fc domain of human IgG1 (ALPROLIX®, 2014; Durocher & Butler, 2009; McCue et al., 2014; Peters et al., 2010). An essential PTM for FIX activity is γ-carboxylation of the first 12 glutamic acid residues in the Gla domain by vitamin K–dependent γ-glutamyl carboxylase. This modification facilitates binding of FIX to phospholipid membranes. HEK293 cells have been reported to have a greater capacity for γ-carboxylation than CHO cells (Berkner, 1993). Furthermore, FVIII contains six potential tyrosine sulfation sites, which are vital for FVIII functionality and binding to von Willebrand factor. FVIII expressed from human cell lines has been reported to be fully sulfated (Kannicht et al., 2013; Peters et al., 2013).
The use of a human cell line for replacement coagulation factors, such as rFVIIIFc and rFIXFc, may result in reduced immunogenicity relative to non-human mammalian cell lines, as α-gal and NGNA glycan moieties are absent from these manufactured protein products (Bosques et al., 2010; McCue et al., 2014, 2015; Noguchi et al., 1995). However, it should be noted that several recombinant clotting factor products produced in non-human mammalian cell lines have been used successfully for many years. The development of inhibitors (neutralizing antibodies) against replacement clotting factors occurs in ∼30% of people with severe hemophilia A and 5% of those with severe hemophilia B. The causative F8 or F9 gene mutation plays a pivotal role in inhibitor development in hemophilia A and B, respectively, with large or complete deletions, non-sense mutations or inversions (e.g. intron 22 inversion in the F8 gene) being the most commonly associated mutations (Franchini & Mannucci, 2011). The impact of PTMs on inhibitor development is unknown, and will need further research. Importantly, none of the previously treated people with hemophilia in the phase 1/2a or phase 3 clinical studies developed inhibitors to the rFVIIIFc and rFIXFc fusion products (Mahlangu et al., 2014; Powell et al., 2012, 2013; Shapiro et al., 2012).
Human-cl rhFVIII (NUWIQ®), an additional factor VIII replacement product for the management of hemophilia A, is being produced in the HEK293-F cell line. Like HEK293-H cells, HEK293-F cells are a derivation of HEK293 cells that have been pre-adapted for growth in serum-free culture medium (Casademunt et al., 2012). Human-cl rhFVIII has been approved by the EMA and submitted to the FDA for approval (Octapharma, 2014); this product has been shown to exhibit a similar glycosylation profile to human plasma-derived factor VIII, without α-gal and NGNA (Kannicht et al., 2013).
Glucagon-1-like peptide (GLP-1) Fc fusion protein (dulaglutide) has been approved by the FDA for the treatment of type 2 diabetes mellitus, and is produced using HEK293-EBNA cells (Glaesner et al., 2010; TRULICITY™, 2014). Large clinical trials have demonstrated its superiority over the dipeptidyl peptidase-4 inhibitor antagonist exenatide and its non-inferiority to liraglutide (a GLP-1 agonist), when added on to oral diabetic agents (Dungan et al., 2014; Wysham et al., 2014).
Another human cell line, HT-1080, was produced from a fibrosarcoma with an epithelial-like phenotype (Swiech et al., 2012). With the use of gene activation technology (in which the endogenous DNA promoter is replaced with a more potent type), four approved therapeutic proteins have been produced by Shire (Swiech et al., 2012).
1) Epoetin delta (DYNEPO®) was approved by the EMA in 2002 for the treatment of anemia secondary to chronic renal failure (DYNEPO®, 2007; ELAPRASE®, 2013; REPLAGAL®, 2006; Swiech et al., 2012; VPRIV®, 2013). However, this has been voluntarily withdrawn by the manufacturer for commercial reasons.
2) Iduronate-2-sulfatase (idursulfase; ELAPRASE®) is licensed as enzyme replacement therapy (EMA in 2007 and FDA in 2006) for the treatment of Hunter syndrome (mucopolysaccharidosis II), an X-linked lysosomal storage disorder (ELAPRASE®, 2013).
3) Agalsidase alfa (REPLAGAL®; Shire Human Genetic Therapies, Danderyd, Sweden) was approved by the EMA in 2001 for the treatment of Fabry disease (REPLAGAL®, 2010). Compared with agalsidase beta (FABRAZYME®; Genzyme Therapeutics, Cambridge, MA), which is produced using CHO cells for a similar indication (FABRAZYME®, 2010, 2014), agalsidase alfa has shown similar enzyme kinetics. However, agalsidase alfa demonstrates a lesser uptake into fibroblasts from patients with Fabry disease and also lower concentrations in the kidney, heart and spleen of mice (Lee et al., 2003). A single clinical study has compared the two products; this showed no significant differences for all efficacy outcomes, and there were no differences for the development of antibodies (Vedder et al., 2007).
4) The fourth agent produced in HT-1080 cells, velaglucerase alfa (VPRIV®; Shire Human Genetic Therapies, Lexington, MA), was approved in 2010 (FDA and EMA) for the treatment of type 1 Gaucher disease (DYNEPO®, 2007; ELAPRASE®, 2013; REPLAGAL®, 2006; Swiech et al., 2012; VPRIV®, 2013). Velaglucerase alfa has been compared with two similar products: imiglucerase, produced using CHO cells, and taliglucerase alfa, produced using carrot cells (Ben Turkia et al., 2013; Tekoah et al., 2013).
These products have diverse glycan profiles and the studies have generally shown comparable uptake into macrophages, in vitro enzymatic activity, stability, organ distribution and efficacy (Ben Turkia et al., 2013; Tekoah et al., 2013). However, neutralizing antibodies to imiglucerase were noted in 24% of patients, which had an impact on enzyme activity. It was noted that various factors, such as the production cell line and glycosylation, may be responsible for the difference in immunogenicity, however, the specificity of the anti-imiglucerase antibodies was not stated (Ben Turkia et al., 2013).
Notably, studies that evaluated epoetin delta produced in HT-1080 cells demonstrated differences in glycosylation compared with erythropoietin produced in CHO cells, including a lack of NGNA in the proteins (Butler & Spearman, 2014; Llop et al., 2008; Shahrokh et al., 2011). However, there were additional overlapping isoforms present in endogenous human erythropoietin isolated from urine and serum relative to epoetin delta that could not be accounted for by sialic residues alone.
Human cell lines used in the expression of proteins in clinical and preclinical development
Human cell lines have been extensively utilized for the production of products that are currently in clinical development. In addition, human cell lines are a frequently used expression system for biomedical research due to their production of human PTMs and high productivity. As productivity may vary across clonal isolates, it is important to screen for those clones with the highest yield of the therapeutic protein (Berkner, 1993).
The PER.C6 cell line was created from human embryonic retinal cells, immortalized via transfection with the adenovirus E1 gene (Havenga et al., 2008). This system was originally developed for the production of human adenovirus vectors for use in vaccine development and gene therapy (Butler & Spearman, 2014). An investment was made in this cell line in order to develop a human expression system, and now an advantage of PER.C6 is its ability to produce a high level of protein when used in the production of human IgG (Jones et al., 2003). However, this does not require amplification of the incorporated gene (Jones et al., 2003). Currently, a variety of products utilizing the PER.C6 cell line are in phase 1 or 2 clinical trials (Durocher & Butler, 2009), including the MOR103 mAb, a human IgG antibody against granulocyte macrophage colony-stimulating factor, and CL184, an antibody against the rabies virus (Nagarajan et al., 2014).
MOR103 is in clinical development for the treatment of rheumatoid arthritis and multiple sclerosis. In a phase 1b/2a, randomized, placebo-controlled study, MOR103 was active in patients with moderately severe rheumatoid arthritis; a small number of patients developed anti-MOR103 antibodies (Behrens et al., 2014). CL184 is a combination of two mAbs, human IgG1(λ) and human IgG1(κ) (Bakker et al., 2005). In a phase 1 clinical study, it demonstrated a favorable safety profile and rapid development of rabies virus neutralizing activity, while there was no evidence to suggest the development of human anti-human antibodies (Bakker et al., 2008). CL184 has been granted FDA fast-track approval status.
Two additional cell lines are utilized by products currently in preclinical development. The CAP cell line is derived from human amniocytes obtained through amniocentesis; these cells are immortalized through an adenovirus type 5 E1 gene (Schiedner et al., 2008; Swiech et al., 2011). In addition to the ability to produce human PTMs, the primary advantage of this cell is the potential for high protein yields (Schiedner et al., 2008).
The HKB-11 cell line was created through polyethylene glycol fusion of HEK293-S and a human B-cell line (modified Burkitt lymphoma cells) (Cho et al., 2003; Durocher & Butler, 2009; Picanco-Castro et al., 2013). The advantages of this cell line include high-level protein production without the formation of aggregates, which can be a problem in other human cell lines (Picanco-Castro et al., 2013). Notably, HKB-11 has demonstrated increased expression of human FVIII compared with expression in HEK293 and BHK21 (Mei et al., 2006). Similar to other human cell lines, it has been shown to produce human glycosylation patterns including α (2,3) and α (2,6) sialic acid linkages (Picanco-Castro et al., 2013). HKB-11 has been used to produce a recombinant factor VIII protein and tissue factor (Cho et al., 2003).
A more recently developed cell line, HuH-7, originates from a human hepatocellular carcinoma (Enjolras et al., 2012). A recent study has shown that the HuH-7-CD4 clone is capable of producing recombinant human factor IX with a human glycosylation profile. PTM profiles (e.g. glycosylation, sialylation, phosphorylation and sulfation) were similar to plasma-derived and recombinant factor IX (rFIX), and were improved relative to rFIX produced in CHO cells (Enjolras et al., 2012). More recently, the HuH-7 cell line has been used to produce mutant forms of rFIX that have improved binding affinity for activated FVIII, and also demonstrated enhanced clotting activity in mice (Perot et al., 2015).
Perceptions of risks versus benefits of using human cell lines
The human-specific glycosylation pattern of the PTMs produced by human cell lines offer several advantages compared with those produced in animal cell lines. Although other mammalian cells can produce similar PTMs to human cells, most also produce α-gal and NGNA, PTMs that are not present in the structure of human proteins (Ghaderi et al., 2012). Patterns of post-translational glycosylation are known to affect protein yield, bioactivity, and clearance (Ghaderi et al., 2010). In addition, antibodies to NGNA have been widely reported to occur in humans (Chung et al., 2008; Ghaderi et al., 2012). One study utilizing an NGNA knockout mouse model demonstrated increased immunogenicity of cetuximab due to anti-NGNA antibodies (Ghaderi et al., 2010). In addition, in patients receiving the mAb cetuximab for the treatment of colorectal or head and neck cancers, the majority of severe hypersensitivity reactions observed in clinical trials were associated with pre-existing IgE antibodies against α-gal (Chung et al., 2008; Ghaderi et al., 2012). Such antibodies may alter the efficacy or immunogenicity of proteins with the presence of non-human glycan structures. Thus, human cell lines can serve as a valuable niche expression system for biotherapeutic proteins that require human PTMs. A theoretical concern with the use of human cell lines is an increased risk of transfer of human adventitious agents, given the lack of a species barrier (Swiech et al., 2012). However, current manufacturing technologies, typically inclusive of multiple viral inactivation or clearance steps, such as nanofiltration, have largely mitigated this concern and may provide more effective viral clearance than has been observed in CHO cells (Kelley et al., 2010; McCue et al., 2014, 2015).
Future perspectives
Production of biotherapeutic proteins in human cell lines is expanding, with several products currently approved for clinical use and others in clinical development in different therapeutic areas. Advantages of human expression systems include achieving equal productivity to other mammalian cell lines and the production of proteins that lack potentially immunogenic, non-human PTMs (most notably α-gal and NGNA). In the future, with additional research investments and a continuation of the technologic advances that have already led to improvements in the use of human cell lines for protein manufacture, human cell lines will be further optimized, more sophisticated product collection strategies will be developed, and these cell lines may become one of the preferred platforms for protein biotherapeutic production.
All brand names are trademarks of their respective owners.
Declaration of interest
Editorial support for the writing of this manuscript was provided by Melissa Yuan, MD, of MedErgy, and was funded by Biogen. All authors are employees of and hold equity interest in Biogen.
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The AntiGMOParty.com is the only political Party to take an stand against food derived from GMO that are harmful to human beings and animal consumption. http://AntiGMOParty.com/
The AntiGMOParty.com was created for consumers. We believe that everyone has the right to an informed choice about what they eat. Many people are concerned about the potential health risks of products made using the relatively new technology of genetic modification (to learn more, please see our About GMOs page). The Non-GMO Project respects your right to know what’s in the food you’re eating and the products you’re buying.
That’s why we have created the world first Political Party for GMO avoidance. The AntiGMOParty.com the only independent, third party, Party that officially has backed a platform to ban GMO Food for human consumption.
By supporting participating members who run for office that are enrolled in the AntiGMOParty, you become an essential part of the collaborative effort to ensure that there are non-GMO choices now and in the future.
Some of the of the best citizen investigators that I know have put together and compiled one of the best detailed independent investigations Iv seen. They have compiled 100+ clips regarding anomalies about the April 15th 2013 Boston Bombing this is some of the most powerful stuff Iv seen in the search for truth in this independent investigation about the Bombing Boston.
“Knowing the lie travels around the world before the truth can tie it’s shoelaces, I decided to share again a year later after citizen investigators unearthed the truth”. John Etter
John Etter“Plasma Burn” did a very good job exposing Jeff the other day on his YouTube channel.
Now second on my list of clips:
Boston WEAK – Bauman Fraudster…See More
As Iv been surfing around for news feeds on the radio, my Facebook and twitter feeds I have noticed the hot topic trending on the News feeds are about Volcano’s, Earthquakes, and Tsunami’s! A growing number of scientists are starting to worry that a magnetic pole shift, seems to be underway, is the real culprit behind climate change. The magnetic north pole is currently shifting at a faster rate than at any time in human history — almost 40 miles a year.
5 Jesus said to them: “Watch out that no one deceives you.6 Many will come in my name, claiming, ‘I am he,’ and will deceive many.7 When you hear of wars and rumors of wars, do not be alarmed. Such things must happen, but the end is still to come.8 Nation will rise against nation, and kingdom against kingdom. There will be earthquakes in various places, and famines. These are the beginning of birth pains. http://www.biblegateway.com/passage/?search=Mark+13
This has got me to thinking about the Natural events that could happen close to home such as the big one the Yellowstone super-volcano that lies beneath Yellowstone National Park in Wyoming that Threatens Two Thirds of USA. For 640,000, year now it has been dormant but when she blows her top it will spew out enough ash and magma to change the world as we know it. Scientists have calculated that the global risk posed by a super-volcanic eruption between five and ten times greater than the probability of being struck by a giant asteroid.
A super-eruption at Yellowstone would be far more devastating for the world than the eruptions at Tambora in 1815, Krakatoa in 1883 and Pinatubo in 1991 which all caused global climate disturbances for several years after the event. Super-eruptions are hundreds of times larger than thebiggest volcanic explosions of recorded history and their effects on the global climate are much more severe, said Professor Stephen Self, a vulcanologist at the Open University. http://www.rense.com/general63/yellowstonesslumbering.htm
“An area the size of North America can be devastated and pronounced deterioration of global climate would be expected for a few years following the eruption,” Professor Self explained. “They could result in the devastation of world agriculture, severe disruption of food supplies and mass starvation. These effects could be sufficiently severe to threaten the fabric of civilization.”
Personally if I lived with in the ground Zero area or in the fallout zone of a super-volcano, I would be moving my family away from that area of the world ASAP! I don’t want to be the doom and gloom guy but I sure would not feel safe knowing that at anytime my family would be wiped of the face of the earth. Sure it has not had an eruption in some 640,000, years but its one day closer everyday. I sure hope if your reading this that you think about moving from those area’s before a big one hits! And if you need a good moving company my good friend Dan Proper is the best and has a dependable moving business called The-Proper-Moving-Company
Pole shift hypothesis
The cataclysmic pole shift hypothesis suggests that there have been geologically rapid shifts in the relative positions of the modern-day geographic locations of the poles and the axis of rotation of … Wikipedia
Related topics
Pole shift hypotheses are not the same as geomagnetic reversal, the periodic reversal of the Earth’s magnetic field (effectively switching the north and south magnetic poles). Wikipedia
The cataclysmic pole shift hypothesis suggests that there have been geologically rapid shifts in the relative positions of the modern-day geographic locations of the poles and the axis of rotation of the Earth, creating calamities such as floods and tectonic events. Wikipedia
On Sunday, the worst earthquake in about 30 years rattled the Yellowstone supervolcano. Overall, there have been at least 25 significant earthquakes at Yellowstone National Park since Thursday, but it is the 4.8 earthquake that has many observers extremely worried.
Image: Yellowstone National Park (Wiki Commons).
Could such a large earthquake be a sign that the Yellowstone supervolcano is starting to roar to life after all this time? And if it does erupt, what would that mean for the rest of the country? As you will see below, a full-blown eruption at Yellowstone would be absolutely catastrophic. It is estimated that such an eruption could dump a 10 foot deep layer of volcanic ash up to 1,000 miles away and render much of the nation uninhabitable for years to come. In essence, it would instantly bring the United States to its knees.
It is true that it is normal for Yellowstone to experience up to 3,000 earthquakes a year. But most of those earthquakes are extremely small and nothing to worry about.
But the 4.8 earthquake that struck on Sunday is definitely raising eyebrows – especially considering what else has been going on at Yellowstone lately.
For example, the scientists that monitor Yellowstone are telling us that the area where the earthquake was centered has been experiencing “ground uplift” in recent months…
A University of Utah release said that the quake area had experienced a “ground uplift” since August and that “seismicity in the general region of the uplift has been elevated for several months.”
I don’t know about you, but the fact that the largest volcano in the U.S. by far has been experiencing “ground uplift” is not very comforting to me.
And there have been reports of strange animal behavior around Yellowstone as well. For example, the following YouTube video of numerous bison literally running away from Yellowstone has gone viral…
That video was captured during the month of March well before the 4.8 earthquake happened.
Could it be possible that those bison sensed that something was coming?
The danger posed by Yellowstone should not be underestimated.
This is something that I have written about before, but since then scientists have discovered that the Yellowstone supervolcano is actually two and a half times larger than they previously believed it to be…
Late last year a new study into the enormous super volcano found the underground magma chamber to be 2.5 times larger than previously thought — a cavern spanning some 90km by 30km and capable of holding 300 billion cubic kilometres of molten rock.
If the sleeping giant were to wake, the outflow of lava, ash and smoke would devastate the United States and affect the entire world.
A full-blown eruption at Yellowstone would be unlike anything that any of us have ever seen before. The following YouTube video attempts to portray what would happen to areas within a few hundred miles of Yellowstone…
But of course the devastation would not just be limited to the northwest part of the country. The following are some more facts about Yellowstone that I compiled for a previous article…
#1 A full-scale eruption of Yellowstone could be up to 1,000 time more powerful than the eruption of Mount St. Helens in 1980.
#3 The next eruption of Yellowstone seems to be getting closer with each passing year. Since 2004, some areas of Yellowstone National Park have risen by as much as 10 inches.
#4 There are approximately 3,000 earthquakes in the Yellowstone area every single year.
#5 In the event of a full-scale eruption of Yellowstone, virtually the entire northwest United States will be completely destroyed.
#6 A massive eruption of Yellowstone would mean that just about everything within a 100 mile radius of Yellowstone would be immediately killed.
#7 A full-scale eruption of Yellowstone could also potentially dump a layer of volcanic ash that is at least 10 feet deep up to 1,000 miles away.
#8 A full-scale eruption of Yellowstone would cover virtually the entire midwest United States with volcanic ash. Food production in America would be almost totally wiped out.
#9 The “volcanic winter” that a massive Yellowstone eruption would cause would radically cool the planet. Some scientists believe that global temperatures would decline by up to 20 degrees.
#10 America would never be the same again after a massive Yellowstone eruption. Some scientists believe that a full eruption by Yellowstone would render two-thirds of the United States completely uninhabitable.
#11 Scientists tell us that it is not a matter of “if” Yellowstone will erupt but rather “when” the next inevitable eruption will take place.
In essence, a Yellowstone eruption would be on the same level as a Carrington event. Either one would fundamentally change life in the United States in a single day.
Personally, I certainly hope that we do not see an eruption at Yellowstone any time soon. And actually, I am much more concerned about the possibility of an eruption at other volcanoes in the northwest such as Mt. Hood and Mt. Rainier.
But if the ground keeps rising rapidly at Yellowstone and earthquakes like the one that struck on Sunday keep on happening, then it would be very foolish for us to ignore the warning signs.
And of course you shouldn’t expect the government to warn you about the potential threat of a Yellowstone eruption until the very last moment. Generally speaking, the government is much more concerned about “keeping people calm” than it is about telling us the truth.
We seem to have moved into a time of increased seismic activity all over North and South America. In such an environment, it would not be wise to say that an eruption at Yellowstone “can’t happen”.
The truth is that an eruption at Yellowstone could happen at any moment. Let us just hope that we are all out of the way when it does.
This article was posted: Tuesday, April 1, 2014 at 5:15 am
Peru Volcano Comes Back to Life Causes Evacuations
A volcano in Peru that has not blown its top in four decades has spewed more ash skyward, after authorities evacuated villagers to avoid Ubinas’s wrath reports SBS.
The volcano in southwestern Peru blasted back to life causing about 60 villagers from Querapi, near its base, to be relocated Saturday, Ubinas town mayor Pascual Coaquira said.
A disaster can be ostensively defined as any tragic event stemming from events such as earthquakes, floods, catastrophic accidents, fires, or explosions. Wikipedia
Beta: Measuring search interest in topics is a beta feature which quickly provides accurate measurements of overall search interest. To measure search interest for a specific query, select the “search term” option.
The Destruction of the Temple and Signs of the End Times
13 As Jesus was leaving the temple, one of his disciples said to him, “Look, Teacher! What massive stones! What magnificent buildings!”
2 “Do you see all these great buildings?” replied Jesus. “Not one stone here will be left on another; every one will be thrown down.”
3 As Jesus was sitting on the Mount of Olives opposite the temple, Peter, James, John and Andrew asked him privately,4 “Tell us, when will these things happen? And what will be the sign that they are all about to be fulfilled?”
5 Jesus said to them: “Watch out that no one deceives you.6 Many will come in my name, claiming, ‘I am he,’ and will deceive many.7 When you hear of wars and rumors of wars, do not be alarmed. Such things must happen, but the end is still to come.8 Nation will rise against nation, and kingdom against kingdom. There will be earthquakes in various places, and famines. These are the beginning of birth pains.
9 “You must be on your guard. You will be handed over to the local councils and flogged in the synagogues. On account of me you will stand before governors and kings as witnesses to them.10 And the gospel must first be preached to all nations.11 Whenever you are arrested and brought to trial, do not worry beforehand about what to say. Just say whatever is given you at the time, for it is not you speaking, but the Holy Spirit.
12 “Brother will betray brother to death, and a father his child. Children will rebel against their parents and have them put to death.13 Everyone will hate you because of me, but the one who stands firm to the end will be saved.
14 “When you see ‘the abomination that causes desolation’[a] standing where it[b] does not belong—let the reader understand—then let those who are in Judea flee to the mountains.15 Let no one on the housetop go down or enter the house to take anything out.16 Let no one in the field go back to get their cloak.17 How dreadful it will be in those days for pregnant women and nursing mothers!18 Pray that this will not take place in winter,19 because those will be days of distress unequaled from the beginning, when God created the world, until now—and never to be equaled again.
20 “If the Lord had not cut short those days, no one would survive. But for the sake of the elect, whom he has chosen, he has shortened them.21 At that time if anyone says to you, ‘Look, here is the Messiah!’ or, ‘Look, there he is!’ do not believe it.22 For false messiahs and false prophets will appear and perform signs and wonders to deceive, if possible, even the elect.23 So be on your guard; I have told you everything ahead of time.
24 “But in those days, following that distress,
“‘the sun will be darkened, and the moon will not give its light; 25 the stars will fall from the sky, and the heavenly bodies will be shaken.’[c]
26 “At that time people will see the Son of Man coming in clouds with great power and glory.27 And he will send his angels and gather his elect from the four winds, from the ends of the earth to the ends of the heavens.
28 “Now learn this lesson from the fig tree: As soon as its twigs get tender and its leaves come out, you know that summer is near.29 Even so, when you see these things happening, you know that it[d] is near, right at the door.30 Truly I tell you, this generation will certainly not pass away until all these things have happened.31 Heaven and earth will pass away, but my words will never pass away.
The Day and Hour Unknown
32 “But about that day or hour no one knows, not even the angels in heaven, nor the Son, but only the Father.33 Be on guard! Be alert[e]! You do not know when that time will come.34 It’s like a man going away: He leaves his house and puts his servants in charge, each with their assigned task, and tells the one at the door to keep watch.
35 “Therefore keep watch because you do not know when the owner of the house will come back—whether in the evening, or at midnight, or when the rooster crows, or at dawn.36 If he comes suddenly, do not let him find you sleeping.37 What I say to you, I say to everyone: ‘Watch!’”
One group after another is denouncing the genetically modified poison on grocery store shelves, adding to the chorus of voices demanding real untainted food.
“NFC was very proud to introduce the first “Natural Only” kosher supervision,” said NFC Director Rabbi Reuven Flamer. “It’s a logical application of our principle, ‘Start Naturally. Stay that Way.’ Therefore, the Natural Apple K cannot be placed on a product that contains GMOs,” Flamer explained.
“While according to the strict letter of Kosher food law a GMO food ingredient is not prohibited, in our view it is not natural. Additionally, there is a Torah (religious)-based law to ‘guard your health’. GMOs are the number-one growing concern among health-conscious consumers and for businesses in the natural and organic food market, as well as in the conventional food industry,” said Rabbi Flamer.
“Recent studies show that GMOs may cause various kinds of health problems from digestive disturbances to food allergies, and that GMOs require more herbicides, which is really the opposite reason why GMOs were touted to be so environmentally helpful in the first place,” Rabbi Flamer added. “For all of the many reasons that GMOs raise a red flag, consumers simply don’t want them in their foods, and our clients want to accommodate their customers.”
Over the next 12 months, the company will phase out the certification of any product that contains GMO ingredients, and will no longer accept applications for certification of products that contain GMOs.
NFC has numerous natural food certification programs, including USDA Organic certification, Kosher certification (under the “Apple K” label), Vegan certification, and Gluten Guard, a gluten-free assurance program.
Each product submitted by a manufacturer for approval is carefully analyzed. The press release explains the process for all of the certification categories. ”The process may include, but is not limited to, a request and review of the ingredient deck including country of origin andcertificate of analysis, product testing, as well as inspection of manufacturing facilities.”
Whether or not your faith requires you to follow the Kosher food laws, this news should be celebrated by anyone who hopes to see the demise of Monsanto and the products created by their mad scientists. While countries across the world are banning GMOs, the wheels are moving slowly in North America to even have GMOs labeled so that consumers can make an informed decision. To have a large demographic refuse to allow genetically modified material in their food is yet another volley against the corruption that is evident in the unholy alliance of the FDA and Monsanto.
Many of us are preparing for the end of times; because as we all know it it is a matter of when not if. While many of us out there are preparing for the complete collapse of the world as we know it not many of us are preparing for a Financial Doomsday which will probably happen sooner than nature going haywire on us. So how exactly do we prepare for a financial Doomsday/ Economic collapse. Through intensive research I have found way that I think will help anyone survive and Economic/ Financial Doomsday.
This one is a bit of a obvious one but I have to put it out there. A shockingly high number of American families are operating without any kind of financial cushion whatsoever.
-According to a Harris Interactive survey taken in 2010, 77 percent of all Americans are living paycheck to paycheck.
-According to one recent survey, one out of every three Americans would not be able to make a mortgage or rent payment next month if they suddenly lost their current job.
This is one reason why so many Americans have lost their homes and why so many Americans have fallen below the poverty level in recent years. They simply had no cushion.
Last year, 2.6 million more Americans dropped into poverty. That was the largest increase that we have seen since the U.S. government began keeping statistics on this back in 1959.
Don’t let this happen to you. At a minimum, everyone out there should have a cushion that will cover at least 6 months worth of expenses. Preferably, you should have a cushion that will last you at least a year or longer. I know that sounds like a lot but trust me you will love having this around in case something does happen to you and your family.
The easiest way I found to do this is I used to buy a cup of coffee from Quick Trip every morning, it would cost me with tax 2.02 (it was their medium size I think if I remember correctly.). Then one day I really got to thinking how much could I save if I stopped drinking coffee well at least buying it from QT every morning. So I did simple math 2.02×7 is $14.14 a week or $56.56 a month about $678.72 a year! And that was just a cup of coffee I of course cut out eating out, junk food for the most part and I am now saving well over $200 a month and putting all of that into savings. That is just one example of how I did it. I am sure there are many ways out there of saving money I have probably not even thought up of yet and I am talking about small things not drastic. Not to mention the cut of junk food has made me happier and healthier.
Investments
Now I want to be clear with this. This is not your typical investment that I am about to talk about. But rather one more suited towards the fall of an economy and essentially the dollar. The most obvious investment you can make is to buy gold and silver. I think every person in this country is doing that at this point. While that will hold value and its all and good, you can not eat gold and silver. At one point people will want food rather than gold. So a gold and silver investment while good should not be the only investment.
Once the economy fails and everyone is running around trying to figure out what to do and items and resources become scares, everything will start becoming more localized and we will probably move back to a barter system. With that said it will be good to prepare for a barter type of system just in case it ends of that way and even if it doesn’t it will not harm you in any way. Here is what I think you will need in a barter type of system.
1. Water, The single most important item any one person can have. Drinkable water is important to life. While I would stock up a lot on this you can also invest in rain catching gear, water purifying system and other things to make undrinkable water drinkable.
2. Salt, stock up on this and start now. This one little thing that people take for granted everyday will be essential to your survival and a good thing to have in a barter system. Salt is one of the most important thing a person can have in their stock pile period. It is essential for the human body, it makes food yummy, and it can preserve food as well.
4. Live stock, food is one of those thing you have to have. While you can hunt if you do not have live stock which is a good way to get meat and other foods live stock is more beneficial. It is there when you need it you don’t have to go out and find it hopefully, you can trade it for other animals goods or services, and depending on the animal it will have many uses.
5. Other currency, while it is good to have food water etc it is also a good idea to have some form of other currency available in case you can some how assuming things are not that bad go to another country or that their currency is not completely useless. This is something I would keep around, because I do not like to assume anything like all currency will be useless and you will be trapped in this country. You just never know what life will throw at you and where you might find yourself. Therefore, I think that having some other currency like Euros, Yen, and the Australian dollar to name a few might be useful.
6. Land, I believe that land is one of those things that over time will increase in value even more so than gold. Because scientist have found a way to make artificial gold believe it or not, but they have yet to find a way without lasing our water supply it will grow in value tremendously. Not to mention land is very useful to have because you can grow food on it have life stock and the uses for land are almost limitless. Not to mention if the land has a natural aquifer.
Like I said not your normal investment portfolio. I could spend so much more time on this but these are the 6 things I think will help you if an economy collapses.
Pay off any debt
Now this one may be a bit tricky to do since many people are having trouble meeting ends meet. But by all means try to pay off any and all debts you may have. I am going to take a guess that banks will want money owed to them if the economy starts failing to keep themselves afloat. Just because the economy fails doesn’t mean our money will be worthless. The great depression is a perfect example of that. Money was not all that worthless still had value just not as much as it does now. This one again may be hard but I know from personal experience but it has to be done. Many things can happen if the American economy can fail don’t be caught off guard pay off any dept you may have. I will have a completely separate blog post about this coming up soon. As I have said I was in that boat I have searched for solutions and I have found it through trial and error. I have given the advice to my friends and it worked great for them. So I will dedicate a whole blog post just to that.
Learn a Skill
Learning a skill is good even if the economy is still going strong. But a carpenter, welder, handyman can be worth their own weight in gold. Knowing how to build and work with your hands will be skills that if the economy goes you can use to barter with. It can be almost any skill just learn one or a trade and master it. I would say master one than have 1000 that you barely know what you are doing.
These are some of things I would recommend. There is so much when it comes to this that I could make this blog post endless, yet I do have to end it at some point. Just remember what can go wrong will go wrong. Do not assume anything prepare for everything. It cant hurt to be ready it can only hurt not to be ready. As we have seen time and time again it is a matter of when not if. If you have any ideas or comments please feel free to post them and let me know how I did or if I missed something important.
Documentary on Stanislaw R. Burzynski’s revolutionary cancer cure treatment based on his discovery on the mechanics of cancer, which lead him to the creation of the Antineoplaston Therapy. Dr. Burzynski’s Therapy has successfully cured thousands of terminal cancer patients for the last 30 years and has demonstrated to be 3 to 5 times more effective than the conventional chemotherapy and radiation treatments.
In spite of the success of his therapy, he has faced the prosecution of big pharma and the FDA which has tried to stop his therapy from spreading in the United States.
I recently started to use Sovereign Silver and I can say that I have noticed a positive difference. I would call it Natures Viagra or energy supplement! I also tried using it when I was sick with a sour throat and was better with in days not weeks! So I can say with my personal experience Yes to the Good for you in a short term! I did not use the recommended dose I would use far less. I used 1/2 a tea spoon twice a day, once in the morning and once at night before bed and that would be about it!
Now for the Bad news about Sovereign Silver it can Kill you if you do not know what your doing. I would say consult your doctor before using Silver products!
Colloidal silver’s proponents will often leave-out the reason why it’s no longer in use by doctors: silver can build-up in your body, make you sick and even kill you. There is a report available online of a 71 year old man who died after taking colloidal silver orally for four months. Here is an excerpt of the report: It seems that some important facts about the 71 year old man who died were left out. My understanding was that he was on pharmaceutical medications that he had just come off of to start taking the colloidal silver. His reactions were consistent for anyone coming off those types of medicines too quickly.
“Department of Clinical Neurological Sciences, University of Western Ontario, London, Ontario, Canada. The authors report a case of a 71-year-old man who developed myoclonic status epilepticus and coma after daily ingestion of colloidal silver for 4 months resulting in high levels of silver in plasma, erythrocytes, and CSF. Despite plasmapheresis, he remained in a persistent vegetative state until his death 5.5 months later. Silver products can cause irreversible neurologic toxicity associated with poor outcome.”.
The Ugly is can turn Blue like a smurf?
One of the most obvious signs of silver-poisoning is that your skin turns a blueish color. Oh, by the way, this change of color is usually permanent. This condition is called Argyria.
There is a Libertarian Party politician in Montana, named Stan Jones, who took homemade colloidal silver, out of fear that theYear 2000 “problem” that had panic-stricken dupes predicting the end of the modern world as we know it, would make modern antibiotics unavailable. So, he self-medicated himself with colloidal silver and it made his skin turn a blue-gray. Here’s a picture I found of him on the Internet. I swear I didn’t doctor it:
What most of the mainstream media conveniently fail to report is that Paul Karason took homemade colloidal silver which he contaminated with salt and drank over a quart a day for years. Despite that, he was given a clean bill of health from Mount Sinai hospital after he had a checkup at the request of the Today Show he appeared on.
Likely no one has consumed more silver, even in the wrong form, than Karason and despite his cosmetic skin condition his clean bill of health stands as a stark refutation to the charges that silver causes harm.
The fact is that millions of people around the world use colloidal silver and yet there are precious few reports of any harm and the blue skin condition known as Argyria is quite rare. In virtually every instance where it is found the cause can be traced to heavy injestion of a product that is not true colloidal silver.
Properly prescribed and administered mainstream drugs, including antibiotics, kill as many as 120,000 people each year by the admission of the American Medical Association.
The main reason that silver fell out of favor was the advent of antibiotics which were patentable and thus much more highly profitable. Likewise, the main reason that colloidal silver is targeted by the trillion dollar a year world pharma empire, mainstream medicine and the media and agencies beholden to them is the threat it represents to the billions of dollars of profits they make from those antibiotics and treatment of conditions colloidal silver remedies.
Calling it a conspiracy would not be inaccurate.
Millions are estimated to use Silver products from top colloidal and ionic silver companies that I am familiar with. Still, where are all the smurfs and where is evidence of all the harm?
There are a grand total of 16 mentions of colloidal silver and argyria in all the voluminous PubMed references. When you remove the homemade ionic silver and the colloidal silver protein that is not really colloidal silver, then you end up with only a handful that might be colloidal silver.
When I tracked down rare reported incidents of Argyria due to ingestion of alleged colloidal silver I have invariably found that it turned out to be contaminated homemade ionic silver, so-called colloidal silver protein (which is particles to large to suspend without protein – and skin has an affinity for protein) or an ionic silver product with far too high ppm silver content.
Bad homemade CS is NOT ‘contaminated ionic silver suspended in protein’. (No-one makes MSP at home). Bad homemade CS is just colloidal silver made in impure water that has been ‘generated’ for too long. Put simply it causes argyria because its way too strong. Paul Karosan and Stan Jones both made that mistake. Paul Karosan continues to do so for some strange reason. (The other famous argyria victim and anti-colloidal silver campaigner, Rosemary Jacobs, actually never drank colloidal silver in her life. She took highly concentrated silver nitrate nose drops (probably around 30,000 ppm) every day for 3 or 4 years when she was about 11. Read her story and she admits this).
The reports at PubMed ranged from bluish fingernail cuticles to one report of death of a 71 year old man, which may or may not have been actual colloidal silver. Just for grins, do a search for “antibiotic side effect deaths”. That returns 675 reports.
Of course Natural News had ads for colloidal silver and colloidal silver makers – the ads are Google ads, which key in on words and phrases in each article the same way Google does with gmail accounts when you send and receive emails. If you went to an article about cancer, you would see ads for cancer treatments.
Now, if you want to say that some products which are labeled as colloidal silver might be dangerous or ineffective, I might agree. Otherwise, it is MY belief that some people make a practice of labeling anything that is not a mainstream approved drug as quackery.
i think the “conspiracy” angle is quite valid. except i’d put it another way. a large industry looking after it’s interests.
There is a general trend to have too much faith in modern medicine. people think its way more advanced then it is. Most people have adopted an attitude that science will save them, but for most people it’s really about healthy lifestyle choices.
There is not much to back up the toxic effects of silver. We use it in silverware, drinking pitchers, jewelry. sure anything can be toxic in huge does.
Iv tried it and found out for myself when I think of all the crap I’ve wasted money on over the years…$35 ain’t much. I really can’t remember the last time a doctor helped me and that wasn’t cheap. More People are killed at hospitals by bad medicine than anything natural.
The reason that deaths from approved drugs are well-known is that such incidents are documented in medical records and there are very real punishments meted-out if anyone tries to cover them up.
The so called Quacks always have an out by simply stating that their product is simply a supplement. The problem with alternative medicine is that most of the aftereffects upon its users are not documented by anyone. Their deaths or complications to their conditions resulting from foregoing standard medical treatment in favor of quacks is merely listed by the resulting condition (e.g. cancer spreads, poisoning, etc) so the effects of quackery aren’t as well-documented, beyond certain articles. Most people who sell these products sure as hell aren’t going to warn anyone about whatever side effects their product’s use might cause. That would be bad for sales and sales are all most company’s really care about.
If I’m cutting into some one’s pocketbook by publishing this, then that’s just too bad.
The bottom line is that silver does work and work very well and there really is very little evidence of harm from properly made and ingested true colloidal silver.
If it did not work, why do you suppose NASA uses it to purify the astronauts drinking water? Its a fact that CS is used to sterilize water in Mir space vehicles and the International space station. http://books.nap.edu/openbook.php?record_id=10942&page=324 There’s perfectly credible science behind this. We are not talking about pyramids and crystals. Or Potters for Peace uses it purify drinking water in third world countries?
Actively Charged As corroborated by several universities, Sovereign Silver contains 96% positively charged silver particles [Ag(n)+], making it at least 34 times more powerful than other brands.
Easily Absorbed Sovereign Silver’s unprecedented particle size of 0.8 nanometers (validated by Transmission Electron Microscopy) allows for easy absorption and excretion from the body.
Less Is More The smaller the particle size, the greater the surface area and the higher the efficiency. That’s why even with a low concentration of 10 ppm, Sovereign Silver is still much more effective than brands which contain up to 500 ppm!
Perfectly Safe Sovereign Silver is formulated to be safe for the whole family. Taken 7 times a day for 70 years, Sovereign Silver still falls below the EPA daily Oral Silver Reference Dose (RfD).
99.999% Pure Sovereign Silver has only two ingredients: pure silver and pharmaceutical grade purified water. It does not contain added salts or proteins that render other silver products less effective. Plus, It is packaged in non leaching glass bottles to guarantee purity throughout it’s shelf life.
For thousands of years, silver has played an essential role in safeguarding human health. In fact, until 1938, colloidal silver silver water the preferred choice of physicians for empowering the immune system and stimulating the body’s innate healing processes*
Today, as more people embrace natural ways to maintain their health and well being, silver is experiencing a resurgence in popularity. And Sovereign Silver is leading the way. By developing technologically advanced refinements in the production of silver colloids, Sovereign Silver Bio Active Silver Hydrosol delivers advantages no other manufacturer can match.
For details call 1-888-328-8840
Made In USA
*These statements have not been evaluated by the Food and Drug Administration. These products are not intended to diagnose, treat, cure or prevent any disease.
Directions
Adults: 1 teaspoon, hold under tongue for 30 seconds, then swallow.
Children 4 years & older: 1/2 teaspoon.
Guidelines:
Maintenance: Once daily.
Immune Building: 3 times daily.
Long Term Immune Support: 5 times daily
Short term immune support: 7 times daily.
*According to the EPA (CASRN7440-22-4) daily Oral Silver Reference Dose (RfD) applied to 10 ppm, one may ingest 178,850 servings safely over 70 years.
Supplement Facts
Serving Size:
1 teaspoon (5ml)
Servings Per Container:
94.5
Silver
50 mcg*
<td* Daily Value not established.
Pharmaceutical Grade Purified Water (USP-NF)
sovereign silver
For more information about the truth about colloidal silver and how mainstream medicine has suppressed alernative and natural healing, see:
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