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Cho s

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The CHO-S is a cell line designed for use in biopharmaceutical research and development. It is a suspension-adapted Chinese Hamster Ovary (CHO) cell line that can be used for the production of recombinant proteins and other biotherapeutics.

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Lab products found in correlation

Cd cho medium, by Thermo Fisher Scientific (5 mentions) Glutamax, by Thermo Fisher Scientific (4 mentions) Penicillin, by Thermo Fisher Scientific (4 mentions) Streptomycin, by Thermo Fisher Scientific (4 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (4 mentions) L glutamine, by Thermo Fisher Scientific (3 mentions) Trypan blue solution, by Nacalai Tesque (2 mentions) Ham s f12, by Fujifilm (2 mentions) Penicillin streptomycin solution, by Nacalai Tesque (2 mentions) Cell counter tc 20, by Bio-Rad (2 mentions) Co2 incubator, by Panasonic (2 mentions) Anti clumping agent, by Thermo Fisher Scientific (2 mentions) Polycarbonate erlenmeyer flask, by Corning (1 mentions) Geneticin g418, by Thermo Fisher Scientific (1 mentions) Zeocin, by Thermo Fisher Scientific (1 mentions) Tc20 automated cell counter, by Bio-Rad (1 mentions) Sodiumpyruvat, by Thermo Fisher Scientific (1 mentions) Methotrexate, by Merck Group (1 mentions) Ht supplement, by Thermo Fisher Scientific (1 mentions) Pluronic f 68, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

CHO-S cells (84 citations) Free style CHO-S cells (11 citations) CHO-S-SFM II (11 citations) CHO-S suspension cells (7 citations) CHO-S-SFM II medium (6 citations) FreeStyle Max CHO-S Expression System (3 citations) Freestyle™ CHO—S (3 citations) CHO-S cell line (3 citations) CHO-S wild type cells (2 citations) FreeStyle CHO-suspension (CHO-S) cells (2 citations)

19 protocols using cho s

1

Cell Culture Conditions for Cytotoxicity Assays

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SEM (55 (link)), Jurkat, CEM, MOLT-16 and Nalm-6 cells (DSMZ) were cultured in RPMI 1640 GlutaMax-I medium (Thermo Fisher Scientific) supplemented with 10% fetal calf serum (FCS; Thermo Fisher Scientific), 100 U/mL penicillin and 100 µg/mL streptomycin (Thermo Fisher Scientific) (R10+). BHK-CD16a (FcγRIIIa V158) cells were cultured in R10+ medium supplemented with 10 μmol/l methotrexate (Sigma-Aldrich) and 500 μg/ml geneticin (Thermo Fisher Scientific) as described before (53 (link), 54 (link), 56 (link)). CHO-CD32a (FcγRIIa H131) cells were cultivated in DMEM medium (Thermo Fisher Scientific) supplemented with 10% FCS, 100 U/ml penicillin and 100 µg/ml streptomycin (57 (link)). Medium was supplemented with 1% NEAA (Thermo Fisher Scientific), 1% sodiumpyruvat (Thermo Fisher Scientific) and 500 µg/ml geneticin. Chinese hamster ovary cells (CHO-S, Thermo Fisher Scientific) were cultured in serum-free CD-CHO medium (Thermo Fisher Scientific) containing 1% HT supplement (Thermo Fisher Scientific) and 2 mM GlutaMax (Thermo Fisher Scientific). After transfection CHO-S cells were cultured in CD OptiCHO medium (Thermo Fisher Scientific) containing 1% HT supplement, 2 mM GlutaMax and 0.1% Pluronic F-68 (Thermo Fisher Scientific).
Gehlert C.L., Rahmati P., Boje A.S., Winterberg D., Krohn S., Theocharis T., Cappuzzello E., Lux A., Nimmerjahn F., Ludwig R.J., Lustig M., Rösner T., Valerius T., Schewe D.M., Kellner C., Klausz K, & Peipp M. (2022). Dual Fc optimization to increase the cytotoxic activity of a CD19-targeting antibody. Frontiers in Immunology, 13, 957874.
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2

Culturing CHO Cells for Suspension

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Chinese hamster ovary cells for suspension culture (CHO-S) were purchased from Gibco, USA. CHO 1-15 cells (CRL-9606) were purchased from ATCC, USA. Ham's F-12 (Wako Pure Chemical Corporation, Japan) with 10% FBS (Thermo Fisher Scientific, USA) and CHO-S-SFM II (Thermo Fisher Scientific, USA) were used for culture of CHO-S and CHO 1-15 cells, respectively. All media were supplemented with penicillin/streptomycin solution (Nacalai Tesque, Japan). Cells were maintained in CO 2 incubator (Panasonic, Japan), at 37 °C with air containing 5% CO 2 . Cell number was measured by Cell Counter TC20 (Bio-Rad, USA) after staining cells with 0.5% trypan blue solution (Nacalai Tesque, Japan).
Ueki M., Tansho N., Sato M., Kanamori H., Ito Y., & Kato Y. (2020). Improved cultivation of CHO cells in bioreactor with reciprocal mixing.
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3

Optimization of TGE in CHO and HEK-293E cells

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Cell lines tested for TGE included CHO-S (Invitrogen), a CHO-S cell line stably expressing Bcl-xL created using the same vector used for the transient expression of Bcl-xL as described below, and the HEK-293E (ATCC) cell line. The HEK-293E cell line is a suspension adapted HEK293 cell line stably expressing the Epstein–Barr virus nuclear antigen (EBNA-1) allowing for episomal replication of ori-P containing plasmids, and has been shown to increase transgene expression [25 (link)]. CHO cells were maintained in SFM4CHO (Hyclone) media supplemented with 8 mmol L-glutamine and 10 ml/L HT supplement while HEK-293E cells were maintained in a 50/50 mixture of SFM4HEK 293 (Hyclone) and FreeStyle 293 (Gibco). These media are here to after referred to as “maintenance medium”. All cultures were grown in a 37°C incubator with 5% CO2 and shaken at 125 rpm either in 125 mL shake flasks or a six-well plate and passaged at a seeding density of 2 × 105 cells/mL every 3-4 days. Viable cell counts were assessed using the Nova Bioprofile flex (Nova Biomedical) or the Guava EasyCyte plus system (Millipore) with the Nexin or viacount assay per the manufacturer's instructions.
Zustiak M.P., Jose L., Xie Y., Zhu J, & Betenbaugh M.J. (2014). Enhanced transient recombinant protein production in CHO cells through the co-transfection of the product gene with Bcl-xL. Biotechnology journal, 9(9), 1164-1174.
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4

Cell Culture Microscopy Imaging Protocol

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Host cell lines included CHO‐S (Invitrogen) and its subclone CHO‐S/4F11, CHO‐K1 (ECACC CCL‐61) and its subclones CHO‐K1/4F10 and 1D9. The subclones were isolated after multiple rounds of cell sorting for increased transient productivity 27. Producer cell lines were CHO‐S‐Humira, producing the human monoclonal antibody Adalimumab and CHO‐K1‐hDAO expressing the copper‐containing human diamine oxidase 28. All cell lines were cultured in CD‐CHO medium (Gibco®), supplemented with 8 mM L‐glutamine (Merck Millipore) and 0.2% Anti‐Clumping Agent (Gibco®) in shaker flasks at 140 rpm, 37°C, 7% CO2. CHO‐K1‐hDAO medium was in addition supplemented with 10 µM CuSO4 (Sigma Aldrich) and 10 µg/mL Blasticidin‐S‐HCl (Life Technologies). Viability was measured by Trypan Blue and better than 95% during all experiments.
For CRM analysis, 1.5 mL cell suspension were centrifuged at 170 × g for 7 min and resuspended in fresh, pre‐warmed growth medium. Cell suspension (15 µL) was applied to a glass microscope slide, covered with a #1.5 glass cover slip (Menzel‐Glaeser, Germany) and sealed with nail polish.
Prats Mateu B., Harreither E., Schosserer M., Puxbaum V., Gludovacz E., Borth N., Gierlinger N, & Grillari J. (2016). Label‐free live cell imaging by Confocal Raman Microscopy identifies CHO host and producer cell lines. Biotechnology Journal, 12(1), 1600037.
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5

Construction of Dual-Expressing CHO Cell Lines

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Construction of Dual-3 and Dual-29 cell lines that express NDST2 and 3OST-1 has been previously reported [15 (link)]. CHO-S® (Thermo Fisher Scientific Inc. Waltham, MA), Dual-3, and Dual-29 cells were maintained in 125 mL polycarbonate Erlenmeyer flasks (Corning, Corning, NY) containing 25 mL of CD CHO medium (Thermo Fisher Scientific) supplemented with 8 mM GlutaMAX™ (Thermo Fisher Scientific) and 15 mL of hypoxanthine/thymidine solution per 500 mL of medium (HT, Corning Life Sciences, Corning, NY). All cells were seeded at 2 × 105 cells/mL and cultured on orbital shakers agitated at 125 rpm in a humidified 37 °C incubator with 5% CO2. In addition, 1 mg/mL of Geneticin (G418, Thermo Fisher Scientific) and 500 mg/mL of Zeocin™ (Thermo Fisher Scientific) were added to the medium for dual expressing cell lines. Viable cell densities were determined by trypan blue exclusion using a Bio-Rad TC20™ automated cell counter.
Baik J.Y., Dahodwala H., Oduah E., Talman L., Gemmill T.R., Gasimli L., Datta P., Yang B., Li G., Zhang F., Li L., Linhardt R.J., Campbell A.M., Gorfien S.F, & Susan T.S. (2015). Optimization of bioprocess conditions improves production of a CHO cell-derived, bioengineered heparin. Biotechnology journal, 10(7), 1067-1081.
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6

Murine MAb 10H4 Production and Characterization

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Murine MAb 10H4 (CPV-2 VP2 specific) was obtained from the National Research Center for Veterinary Medicine, which was produced by the immunization of mice with partially purified new CPV-2a virus (strain CVCC AV298). The host cell lines used in the expression of recombinant antibodies included the ExpiCHO-S (Thermo Scientific, USA) and CHO-S cell lines (Thermo Scientific, USA). The virus of new CPV-2a, new CPV-2b, and CPV-2c were provided by the National Research Center for Veterinary Medicine.
Zhou L., Wu H., Du M., Song H., Huo N., Chen X., Su X., Li W., Wang L., Wang J., Huang B., Tan F, & Tian K. (2022). A canine-derived chimeric antibody with high neutralizing activity against canine parvovirus-2. AMB Express, 12, 76.
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7

Culturing CHO-S and 4T1 Cell Lines

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FreeStyle™ Chinese Hamster Ovary (CHO-S) (Thermo Fisher Scientific, Waltham, MA) and 4T1 (ATCC, Manassas, VA) cells were utilized for in vitro studies. CHO-S cells were passaged in FreeStyle™ CHO Expression Medium (Thermo Fisher Scientific) according to the manufacturer’s recommendations. 4T1 cells are naturally deficient in H-2Kb expression and were grown in RPMI 1640 supplemented with 10% heat-inactivated FBS, 100 U/ml penicillin, 100 μg/ml streptomycin, and 10 mmol/l L-glutamine (all from Thermo Fisher Scientific). All cell lines were maintained in vented flasks at 37 °C with 5% CO2.
Wooster A.L., Anderson T.S, & Lowe D.B. (2018). EXPRESSION AND CHARACTERIZATION OF SOLUBLE EPITOPE-DEFINED MAJOR HISTOCOMPATIBILITY COMPLEX (MHC) FROM STABLE EUKARYOTIC CELL LINES. Journal of immunological methods, 464, 22-30.
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8

Transient Antibody Screening in CHO-S Cells

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The Chinese hamster ovary (CHO-S from Thermo Fisher Scientific, US)-derived cell line CHOEBNALT85 (Icosagen Cell Factory, Estonia) was grown in serum-free chemically defined medium and was used for antibody screening. This cell line expresses EBV EBNA1 protein and mouse polyomavirus large T protein and is specifically designed for the prolonged and high level production of proteins in association with pQMCF vectors (USPTO Patent No: 7,790,446). The cells were transfected using chemical transfection Reagent 007 (Icosagen Cell Factory, Estonia) according to the published protocols [22 (link)]. One microgram of plasmid DNA was transfected in 6-well plate format for analyzing library pools, and approximately 0.2 - 0.5 μg DNA per sample was used in a high-throughput 96-well plate transfection for screening individual clones. Seventy-two hours after transfection, the supernatants were collected for analysis. When necessary, scFv-Fc or human IgG1 concentrations in the samples were determined using FastELISA for Human IgG quantification (RD Biotech, France).
Kivi G., Teesalu K., Parik J., Kontkar E., Ustav M J.r., Noodla L., Ustav M, & Männik A. (2016). HybriFree: a robust and rapid method for the development of monoclonal antibodies from different host species. BMC Biotechnology, 16, 2.
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9

Comparative Analysis of Suspension CHO Cell Lines

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Three suspension CHO Sublines, including CHO K1, CHO/dhfr− and CHO S, were analyzed in this study. The CHO K1 and CHO S were purchased from Thermo Fisher Scientific (Waltham, MA), and CHO/dhfr− was purchased from ATCC (Manassas, VA). The seed culture of CHO K1, CHO S and CHO/dhfr− were maintained in the three basal media of HyClone CDM4CHO (Hyclone Laboratories, Logan, UT), Gibco CD CHO (Life Technologies, Grand Island, NY) and Sigma EX-CELL CHO DHFR- (Sigma-Aldrich, St. Louis, MO), respectively. All the cell culture media were supplemented with 8 mM L-glutamine (final concentration). The sodium hypoxanthine and thymidine supplements were added to the EX-CELL CHO DHFR- medium. The batch cultures were seeded with viable cell density of 0.3×106 cells/mL. The cells were cultivated with triplication in suspension cultures in 125-mL disposable shaker flasks at 37 °C, 5% CO2 and 120 rpm in a humidified incubator (Caron, Marietta, OH).
Xu N., Ma C., Ou J., Sun W.(., Zhou L., Hu H, & Liu X.(. (2017). Comparative Proteomic Analysis of Three Chinese Hamster Ovary (CHO) Host Cells. Biochemical engineering journal, 124, 122-129.
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10

Anti-VEGF Monoclonal Antibody Production in CHO-S Cells

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A CHO-S (Thermo Fisher Scientific, USA, Catalog no: A11364) cell line expressing a 149 kDa anti-VEGF monoclonal antibody was obtained from Research Working Cell Bank (RWCB) of AryoGen Pharmed (Alborz, Iran). Cells were cultured in a chemically defined medium (GE Healthcare, Sweden) supplemented with 6 mM L-glutamine (Lonza, Belgium). Seed culture was prepared in a 500-ml baffled shake flask with an effective volume of 100 ml, incubated at 37°C with 5% CO2 and agitation speed of 75 RPM (Revolution Per Minute) (Infors LTD., UK) with 25 mm orbital diameter. The temperature was down-shifted to 32°C on day 5 of culture.
Ahleboot Z., Khorshidtalab M., Motahari P., Mahboudi R., Arjmand R., Mokarizadeh A, & Maleknia S. (2021). Designing a Strategy for pH Control to Improve CHO Cell Productivity in Bioreactor. Avicenna Journal of Medical Biotechnology, 13(3), 123-130.
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