Some big food and beverage companies, Test Flavorings That Can Mimic Sugar, Salt or MSG including Kraft Foods, Nestle, Coca-Cola and Campbell Soup, are considering adding human genome sequence, chemical product developed by biotechnology company Senomyx that allows them to cut amount of salt or sugar but leaves taste intact; Senomyx’s chemical compounds will not be listed separately on ingredient labels, but will be lumped into broad category–‘artificial flavors’–already found on most 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
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.
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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
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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 …
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Human Dermal Fibroblasts: HDF
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Human Mammary Epithelial Cells: HMEpC
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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
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Human Adipocyte Differentiation Medium
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MCDB 105 Medium
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Human Preadipocytes: HPAd
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Canine Osteoblasts: CnOb
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Human Dermal Fibroblasts: HDF
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Human Chondrocytes: HC
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Human Coronary Artery Endothelial Cells: HCAEC
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Human MesoEndo Endothelial Cell Media
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Rat Aortic Smooth Muscle Cells: RAOSMC
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Human Dermal Fibroblasts: HDF
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Human Aortic Smooth Muscle Cells: HAOSMC
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Human Epidermal Keratinocytes: HEK
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Human Endothelial Cell Growth Medium
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DiI-Ac-LDL Kit
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Human Hair Follicle Dermal Papilla Cells: HFDPC
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Human Aortic Smooth Muscle Cells: HAOSMC
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Endothelial Cell Growth Medium
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Rat Brain Microvascular Endothelial Cells: RBMVEC
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Human Carotid Artery Smooth Muscle Cells: HCtASMC
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Human Fibroblast-Like Synoviocytes: HFLS
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Human Cardiac Fibroblasts: HCF
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Rat Smooth Muscle Cell Media
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Human Smooth Muscle Cell Growth Medium
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Human Fibroblast-Like Synoviocytes: HFLS
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Human Carotid Artery Endothelial Cells: HCtAEC
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Human Fibroblast-Like Synoviocytes: Rheumatoid Arthritis: HFLS-RA
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Bovine Aortic Endothelial Cells: BAOEC
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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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