scholarly journals Rich production media as a platform for CHO cell line development

2020 ◽  
Author(s):  
Yong Jae Kim ◽  
Sang Kyul Han ◽  
Seongtae Yoon ◽  
Chan-Wha Kim
Keyword(s):  
Cell Culture ◽  
Cell Line ◽  
Mammalian Cells ◽  
Culture Media ◽  
Chinese Hamster ◽  
Cell Cloning ◽  
Cell Line Development ◽  
Solid Media ◽  
Semi Solid ◽  
Set Up

Abstract Recent cell culture media for mammalian cells can be abundantly formulated with nutrients supporting production, but such media can be limited to use in host cell culture, transfection, cell cloning, and cell growth under the low cell density conditions. In many cases, appropriate platform media are used for cell line development, and then replaced with rich media for production. In this study, we demonstrate rich chemically defined media for Chinese hamster ovary (CHO) cells that are suitable as basal media both for cell line development and for final production of culture process. Set up for transfection, semi-solid media optimization, mini-pool screening, and single cell cloning media development were performed, and final clones were obtained with higher productivity in fed-batch culture mode using rich formulated media comparing with lean formulated media. Developed methods may remove the requirements for cell adaptation to production media after cell line development, and relieve the clonality issues associated with changing the culture media. Furthermore, established methods have advantages over traditional approaches, including saving resources and decreasing the time and the effort required to optimize the production process.

Formation of hydrogen peroxide in cell culture media by apple polyphenols and its effect on antioxidant biomarkers in the colon cell line HT-29

2009 ◽  
Vol 53 (10) ◽  
pp. 1226-1236 ◽  
Author(s):  
Phillip Bellion ◽  
Melanie Olk ◽  
Frank Will ◽  
Helmut Dietrich ◽  
Matthias Baum ◽  
...  
Keyword(s):  
Hydrogen Peroxide ◽  
Cell Culture ◽  
Cell Line ◽  
Culture Media ◽  
Cell Culture Media ◽  
Colon Cell ◽  
Colon Cell Line ◽  
Ht 29

scholarly journals Is Oxytocin Proper for Cancer Adjuvant Therapy?

Proceedings ◽  
2018 ◽  
Vol 2 (25) ◽  
pp. 1582
Author(s):  
Ali Taghizadehghalehjoughi ◽  
Betul Cicek ◽  
Ahmet Hacimuftuoglu ◽  
Mustaf Gul
Keyword(s):  
Cell Culture ◽  
Cell Viability ◽  
Cell Line ◽  
Culture Media ◽  
Neuroblastoma Cell ◽  
Sympathetic Ganglia ◽  
Control Group ◽  
Human Neuroblastoma ◽  
Lactation Stage ◽  
Cortex Cell

Neuroblastomas are solid tumors and mostly seen in the adrenal medulla and sympathetic ganglia. It is known that neuroblastoma cell proliferation is inhibited by cisplatin and vincristine. The aim of this study was to investigate the effect of oxytocin on cell viability in human neuroblastoma SH-SY5Y cell line and primary cerebral cortex cell culture exposed to cisplatin and vincristine. In this direction, SH-SY5Y cell line and cortex neurons were obtained from the medical pharmacology department, Ataturk University. Both cells were grown in the appropriate cell culture media. Cisplatin (5, 10, 15 μg), vincristine (0.5, 1 and 2 ng) and oxytocin (1 μM) were applied to SH-SY5Y cell line and primary cortex cell culture for 24 h. MTT and TAC-TOS tests were performed 24 h after the application. As a result of the MTT assay, the combination of cisplatin and vincristine reduced cell viability in both cultures approximately 25% and 22%, respectively, compared to the control group. It appears that oxytocin increases neuroblastoma and cortex neuron viability, 112% and 95%, respectively. In this relation, we need to investigate why oxytocin increases cell viability and what are the possible implications in women in lactation stage.

scholarly journals Establishment of fast-growing serum-free immortalised cells from Chinese hamster lung tissues for biopharmaceutical production

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Noriko Yamano-Adachi ◽  
Rintaro Arishima ◽  
Sukwattananipaat Puriwat ◽  
Takeshi Omasa
Keyword(s):  
Cell Line ◽  
Cho Cells ◽  
Mammalian Cells ◽  
High Efficiency ◽  
Chinese Hamster ◽  
Serum Free ◽  
Sterility Testing ◽  
Fast Growing ◽  
Defined Culture

Abstract Chinese hamster (Cricetulus griseus) ovary-derived Chinese hamster ovary (CHO) cells are the most commonly used mammalian hosts for the industrial production of recombinant therapeutics because of their ability to fold, assemble, and perform post-translational modifications, such as glycosylation, on proteins. They are also valuable for their ability to grow in serum-free suspension cultures. In this study, we established a cell line derived from lung tissue of Chinese hamsters, named Chinese hamster lung (CHL)-YN cells. The biosafety of CHL-YN cells was confirmed by in vitro sterility testing, mycoplasma detection, and reverse transcriptase assays. One of the key characteristics of CHL-YN cells was their doubling time of 8.1 h in chemically defined culture medium; thus, they proliferate much faster than conventional CHO cells and general mammalian cells. Transgenes could be introduced into CHL-YN cells with high efficiency. Finally, between 50% to > 100% of the amount of glycosylated immunoglobulin G (IgG)1 produced by CHO-K1 cells was produced by CHL-YN cells over a shorter period of time. In summary, fast-growing CHL-YN cells are a unique cell line for producing recombinant proteins.

scholarly journals Interactions of a neuronal cell line (PC12) with laminin, collagen IV, and fibronectin: identification of integrin-related glycoproteins involved in attachment and process outgrowth.

1987 ◽  
Vol 105 (5) ◽  
pp. 2347-2358 ◽  
Author(s):  
K J Tomaselli ◽  
C H Damsky ◽  
L F Reichardt
Keyword(s):  
Cell Surface ◽  
Cell Line ◽  
Pc12 Cells ◽  
Pc12 Cell ◽  
Mammalian Cells ◽  
Chinese Hamster ◽  
Neuronal Cell ◽  
Collagen Iv ◽  
Nervous System Development ◽  
Process Outgrowth

Neuronal responses to extracellular matrix (ECM) constituents are likely to play an important role in nervous system development and regeneration. We have studied the interactions of a neuron-like rat pheochromocytoma cell line, PC12, with ECM protein-coated substrates. Using a quantitative cell attachment assay, PC12 cells were shown to adhere readily to laminin (LN) or collagen IV (Col IV) but poorly to fibronectin (FN). The specificity of attachment to these ECM proteins was demonstrated using ligand-specific antibodies and synthetic peptides. To identify PC12 cell surface proteins that mediate interactions with LN, Col IV, and FN, two different antisera to putative ECM receptors purified from mammalian cells were tested for their effects on PC12 cell adhesion and neuritic process outgrowth. Antibodies to a 140-kD FN receptor heterodimer purified from Chinese hamster ovarian cells (anti-FNR; Brown, P. J., and R. L. Juliano, 1986, J. Cell Biol., 103:1595-1603) inhibited attachment to LN and FN but not to Col IV. Antibodies to an ECM receptor preparation purified from baby hamster kidney fibroblastic cells (anti-ECMR; Knudsen, K. A., P. E. Rao, C. H. Damsky, and C. A. Buck, 1981, Proc. Natl. Acad. Sci. USA., 78:6071-6075) inhibited attachment to LN, FN, and Col IV, but did not prevent attachment to other adhesive substrates. In addition to its effects on adhesion, the anti-ECMR serum inhibited both PC12 cell and sympathetic neuronal process outgrowth on LN substrates. Immunoprecipitation of surface-iodinated or [3H]glucosamine-labeled PC12 cells with either the anti-FNR or anti-ECMR serum identified three prominent cell surface glycoproteins of 120, 140, and 180 kD under nonreducing conditions. The 120-kD glycoprotein, which could be labeled with 32P-orthophosphate and appeared to be noncovalently associated with the 140- and 180-kD proteins, cross reacted with antibodies to the beta-subunit (band 3) of the avian integrin complex, itself a receptor or receptors for the ECM constituents LN, FN, and some collagens.

Effects of Water and Cell Culture Media on the Physicochemical Properties of ZnMgO Nanoparticles and Their Toxicity toward Mammalian Cells

Langmuir ◽  
2014 ◽  
Vol 30 (38) ◽  
pp. 11366-11374 ◽  
Author(s):  
Jasmina Vidic ◽  
Francia Haque ◽  
Jean Michel Guigner ◽  
Aurore Vidy ◽  
Christophe Chevalier ◽  
...  
Keyword(s):  
Cell Culture ◽  
Physicochemical Properties ◽  
Mammalian Cells ◽  
Culture Media ◽  
Cell Culture Media

scholarly journals S-Trap eliminates cell culture media polymeric surfactants for effective proteomic analysis of mammalian cell bioreactor supernatants

2020 ◽  
Author(s):  
Lucia F. Zacchi ◽  
Dinora Roche Recinos ◽  
Ellen Otte ◽  
Campbell Aitken ◽  
Tony Hunt ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Cell Culture ◽  
Sample Preparation ◽  
Mammalian Cell ◽  
Proteomic Analysis ◽  
Mammalian Cells ◽  
Culture Media ◽  
Quality Data ◽  
Polymeric Surfactants ◽  
Cell Culture Media

AbstractProteomic analysis of bioreactor supernatants can inform on cellular metabolic status, viability, and productivity, as well as product quality, which can in turn help optimize bioreactor operation. Incubating mammalian cells in bioreactors requires the addition of polymeric surfactants such as Pluronic F68, which reduce the sheer stress caused by agitation. However, these surfactants are incompatible with mass spectrometry proteomics and must be eliminated during sample preparation. Here, we compared four different sample preparation methods to eliminate polymeric surfactants from filtered bioreactor supernatant samples: organic solvent precipitation; filter-assisted sample preparation (FASP); S-Trap; and single-pot, solid-phase, sample preparation (SP3). We found that SP3 and S-Trap substantially reduced or eliminated the polymer(s), but S-Trap provided the most robust clean-up and highest quality data. Additionally, we observed that SP3 sample preparation of our samples and in other published datasets was associated with partial alkylation of cysteines, which could impact the confidence and robustness of protein identification and quantification. Finally, we observed that several commercial mammalian cell culture media and media supplements also contained polymers with similar mass spectrometry profiles, and we suggest that proteomic analyses in these media will also benefit from the use of S-Trap sample preparation.

scholarly journals A big cheese in biotherapeutics: Lactoyl leucine and isoleucine are bioavailable alternatives for canonical amino acids in cell culture media

2021 ◽  
Author(s):  
Corinna Schmidt ◽  
Maria Wehsling ◽  
Maxime Le Mignon ◽  
Gregor Wille ◽  
Yannick Rey ◽  
...  
Keyword(s):  
Amino Acids ◽  
Cell Culture ◽  
Cho Cells ◽  
Culture Media ◽  
Chinese Hamster ◽  
Batch Processes ◽  
Next Generation ◽  
Cell Culture Media ◽  
Derivatives Of

Increasing demands for protein-based therapeutics such as monoclonal antibodies, fusion proteins, bispecific molecules and antibody fragments require researchers to constantly find innovative solutions. To increase yields and decrease costs of next generation bioprocesses, highly concentrated cell culture media formulations are developed but often limited by the low solubility of amino acids such as tyrosine, cystine, leucine and isoleucine, in particular at physiological pH. This work sought to investigate highly soluble and bioavailable derivatives of leucine and isoleucine that are applicable for fed-batch processes. N-lactoyl-leucine and N-lactoyl-isoleucine sodium salts were tested in cell culture media and proved to be beneficial to increase the overall solubility of cell culture media formulations. These modified amino acids proved to be bioavailable for various Chinese hamster ovary (CHO) cells and were suitable for replacement of canonical amino acids in cell culture feeds. The quality of the final recombinant protein was studied in bioprocesses using the derivatives, and the mechanism of cleavage was investigated in CHO cells. Altogether, both N-lactoyl amino acids represent an advantageous alternative to canonical amino acids to develop highly concentrated cell culture media formulations to support next generation bioprocesses.

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