Fetal bovine serum (fbs)

Manufactured by Atlanta Biologicals

Sourced in United States, Gabon, Switzerland, Holy See (Vatican City State), Canada, Germany, China, Macao, United Kingdom, Ireland

FBS (Fetal Bovine Serum) is a common laboratory reagent used as a supplement in cell culture media. It provides essential growth factors, nutrients, and other components required for the optimal growth and maintenance of a variety of cell types in vitro.

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FBS (fetal bovine serum) (4 citations) Fetal bovine serum(FBS; S11150) (3 citations) Charcoal dextran stripped fetal bovine serum (CD-FBS) (2 citations) Heat Inactivated Fetal Bovine Serum (FBS-HI) (2 citations) Fetal bovine serum/HI-FBS (2 citations)

2 749 protocols using fetal bovine serum (fbs)

Cell Culture Protocols for Cancer Cell Lines

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HCT-116 and DMS-53 cells were obtained from the ATCC (American Type Culture Collection, Manassas, VA). H1299 cells were kindly provided by the Lung SPORE cell line repository at H. Lee Moffitt Cancer Center & Research Institute. HCT-116/δOR cells were previously generated in our lab using pcDNA-δOR15 vector containing a truncated δOR lacking the final 15 C-terminal amino acids. ^{45 (link)} HCT-116 and HCT-116/δOR cells were cultured in DMEM/F-12 (1:1) media containing 365 mg/L L-Glutamine, 2.438 g/L Sodium Bicarbonate (Life Technologies, Gibco), 10% fetal bovine serum (Atlanta Biologicals), 100 units/mL penicillin, and 100 µg/mL streptomycin. H1299 cells were cultured in RPMI-1640 media containing 300 mg/L L-Glutamine (Life Technologies, Invitrogen), 10% fetal bovine serum (Atlanta Biologicals), 100 units/mL penicillin, and 100 µg/mL streptomycin. DMS-53 cells were cultured in RPMI-1640 media containing 300 mg/L L-Glutamine (Life Technologies, Invitrogen) and 10% fetal bovine serum (Atlanta Biologicals). The cells were incubated in 5% CO₂ at 37 °C. Throughout this study, the morphology and growth characteristics of these cells were monitored by microscopy.

Cohen A.S., Patek R., Enkemann S.A., Johnson J.O., Chen T., Toloza E., Vagner J, & Morse D.L. (2015). Delta-Opioid Receptor (δOR) Targeted Near-Infrared Fluorescent Agent for Imaging of Lung Cancer: Synthesis and Evaluation In Vitro and In Vivo. Bioconjugate chemistry, 27(2), 427-438.

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Cell Culture Protocols for Ewing Sarcoma

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Ewing cells TC32, CHLA10, CHLA9, and TC71 were purchased from Children’s Oncology Group (COG) and the EWS502 Ewing sarcoma cell line was a kind gift from Dr. Stephen Lessnick (Nationwide Children’s Hospital, Columbus, OH, USA). IMR90, a primary fibroblast cell line was used as control and purchased from ATCC. TC32, TC71, and EWS502 were grown in RPMI supplemented with 10% Fetal Bovine Serum (Atlanta Biologicals, Flowery Branch, GA, USA), CHLA10 and CHLA9 in IMDM supplemented with 20% Fetal Bovine Serum and IMR90 in DMEM supplemented with 10% Fetal Bovine Serum. Cells were maintained in 37 °C in a humidified atmosphere with 5% CO₂.

Miller H.E., Gorthi A., Bassani N., Lawrence L.A., Iskra B.S, & Bishop A.J. (2020). Reconstruction of Ewing Sarcoma Developmental Context from Mass-Scale Transcriptomics Reveals Characteristics of EWSR1-FLI1 Permissibility. Cancers, 12(4), 948.

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Influenza Virus Propagation in Mammalian Cell Lines

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MDCK cells [obtained from Dr. Daniel Perez] and MDCK WSN HA cells [obtained from Dr. Ryan Langlois] were maintained Minimal Essential Medium [Sigma] supplemented with 10% fetal bovine serum [FBS; Atlanta Biologicals], penicillin [100 IUml⁻¹], and streptomycin [100 μg ml⁻¹; PS; Corning]. A549 WT, A549 Rab11a KO were maintained in Dulbecco’s Modified Essential Medium [Gibco] supplemented with 10% FBS [Atlanta Biologicals], and PS. All cells were cultured at 37°C and 5% CO₂ in a humidified incubator. All cell lines were tested monthly for mycoplasma contamination while in use. The medium for culture of IAV in each cell line [termed virus medium] was prepared by eliminating FBS and supplementing the appropriate medium with 4.3% BSA and PS. Ammonium chloride-containing virus medium was prepared by the addition of HEPES buffer and NH₄Cl at final concentrations of 50 mM and 20 mM, respectively. OPTi-MEM [Gibco] was used as a serum free medium where indicated.

Ganti K., Han J., Manicassamy B, & Lowen A.C. (2021). Rab11a mediates cell-cell spread and reassortment of influenza A virus genomes via tunneling nanotubes. PLoS Pathogens, 17(9), e1009321.

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Cell Culture Protocols for Vero and BSR-T7 Cells

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Vero cells, a spontaneously immortalized cell line isolated from the kidney of an adult female African grivet monkey (RRID:CVCL_0059), were obtained from the American Type Culture Collection (ATCC). Cells were cultured in Dulbecco’s Modified Eagle Medium (DMEM; Thermo Fisher) supplemented with 2% fetal bovine serum (Atlanta Biologicals), 1% GlutaMAX (Thermo Fisher), and 1% penicillin-streptomycin (Thermo Fisher). BSR-T7 cells (RRID:CVCL_RW96), generated by stable T7 RNA polymerase expression in BHK-21 cells, were a kind gift from K.-K. Conzelmann. The parent cell line (RRID: CVCL_1915) was isolated from the kidney of a 1-day old male golden hamster. BSR-T7 cells were cultured in Dulbecco’s Modified Eagle Medium (DMEM; Thermo Fisher) supplemented with 10% fetal bovine serum (Atlanta Biologicals), 1% GlutaMAX (Thermo Fisher), and 1% penicillin-streptomycin (Thermo Fisher). Vero E6, a spontaneously immortalized cell line isolated from the kidney of an adult female African grivet monkey (RRID: CVCL_0574) were obtained from ATCC. Cells were cultured in Dulbecco’s Modified Eagle Medium (DMEM; Thermo Fisher) supplemented with 2% fetal bovine serum (Atlanta Biologicals), 1% GlutaMAX (Thermo Fisher), and 1% penicillin-streptomycin (Thermo Fisher). All cell lines were maintained in a humidified 37°C incubator supplied with 5% CO2. Cell lines were not authenticated following purchase.

Fels J.M., Maurer D.P., Herbert A.S., Wirchnianski A.S., Vergnolle O., Cross R.W., Abelson D.M., Moyer C.L., Mishra A.K., Aguilan J.T., Kuehne A.I., Pauli N.T., Bakken R.R., Nyakatura E.K., Hellert J., Quevedo G., Lobel L., Balinandi S., Lutwama J.J., Zeitlin L., Geisbert T.W., Rey F.A., Sidoli S., McLellan J.S., Lai J.R., Bornholdt Z.A., Dye J.M., Walker L.M, & Chandran K. (2021). Protective neutralizing antibodies from human survivors of Crimean-Congo hemorrhagic fever. Cell, 184(13), 3486-3501.e21.

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Cell Culture Conditions for Multiple Cell Lines

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All cells were incubated at 37 °C with 5% CO₂. Huh7,
HepG2, PH5CH8 and HEK 293 cells were cultured in D MEM (Sigma) containing 10%
heat-inactivated FBS (Atlanta Biologicals) and 1%
penicillin-streptomycin-glutamine (Mediatech). BHK cells were cultured in MEM
(Sigma) containing 10% heat-inactivated FBS (Atlanta Biologicals) and 1%
penicillin-streptomycin-glutamine (Mediatech). CAL-1 cells were grown in RPMI
1640 (Sigma) containing 10% heat-inactivated FBS (Atlanta Biologicals) and 1%
penicillin-streptomycin-glutamine (Mediatech), and supplemented with 2 mM
L-Glutamine, 1 mM sodium pyruvate, 10 mM HEPES, 1x MEM NEAA (Corning).

Schwerk J., Soveg F.W., Ryan A.P., Thomas K.R., Hatfield L.D., Ozarkar S., Forero A., Kell A.M., Roby J.A., So L., Hyde J.L., Gale M J.r., Daugherty M.D, & Savan R. (2019). RNA-binding protein isoforms ZAP-S and ZAP-L have distinct antiviral and immune resolution functions. Nature immunology, 20(12), 1610-1620.

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Cell Culture Optimization and Proteomic Conditions

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We cultured all cells in humidified incubators at 37 °C with 5% CO₂. Adherent U2OS cells (from Cell and Genome Engineering Core, UCSF) were grown to ~90% confluency in DM EM media (Corning) supplemented with 10% FBS (Axenia BioLogix) on 3.5 cm (for optimizing conditions), or 10 cm (for proteomics) dishes. The Jurkat E6.1 T cells (a gift from Dr. Arthur Weiss, UCSF) were maintained in suspension at 0.2e6 to 1e6 cells/mL cell density either in regular RPMI1640 media supplemented with 10% FBS (Atlanta Biologicals) (for optimizing conditions) or in advanced RPMI1640 medium (Life Technologies) supplemented with 2% in-house heat-treated (56 °C for 30 min) FBS (Atlanta Biologicals) and GlutaMAX (for proteomics). AMO1 cells (a gift from Dr. Christoph Driesen, Kantonsspital St. Gallen, Switzerland) in suspension were cultured at 1e6 to 2e6 cells/mL cell density in RPMI 1640 medium supplemented with 1% penicillin/streptomycin (ThermoFisher Scientific) and 20% FBS (Atlanta Biologicals). We confirmed mycoplasma negativity for all cultures prior to experiments (DigitalTest v2.0, biotool.com). Replicate experiments were performed on cells from different passages.

Tuomivaara S.T., Teo C.F., Jan Y.N., Jan L.Y, & Wiita A.P. (2023). SLAPSHOT reveals rapid dynamics of extracellularly exposed proteome in response to calcium-activated plasma membrane phospholipid scrambling. bioRxiv.

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Culturing Human Cell Lines

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Human esophageal squamous epithelial cancer cell line TE-7 was kindly provided by Dr. Hainault (IARC, Lyon, France). These cells were maintained in RPMI medium (Invitrogen) supplemented with 5% FBS (Atlanta Biologicals) and 1% penicillin/streptomycin (Invitrogen). HEK 293T cells and L929 cells were grown in DMEM medium (Invitrogen) supplemented with 10% FBS (Atlanta Biologicals) and 1% penicillin/streptomycin (Invitrogen). Jurkat cells were cultured in RPMI medium (Invitrogen) supplemented with 10% FBS (Atlanta Biologicals) and 1% penicillin/streptomycin (Invitrogen).

Caldwell J.M., Collins M.H., Kemme K.A., Sherrill J.D., Wen T., Rochman M., Stucke E.M., Amin L., Tai H., Putnam P.E., Jiménez-Dalmaroni M.J., Wormald M.R., Porollo A., Abonia J.P, & Rothenberg M. (2017). Cadherin 26 is an alpha integrin-binding epithelial receptor regulated during allergic inflammation. Mucosal immunology, 10(5), 1190-1201.

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Culturing Human Cell Lines

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Breast Cancer Cell Line Maintenance Protocol

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MCF-7 (HER2 positive), MDAMB-231 (TNBC), BT-549 (TNBC), and SK-BR-3 (HER-2 positive) breast cancer cell lines were obtained from ATCC (Manassas, VA, USA). MCF-7, and MDAMB-231 were maintained in DMEM medium (Corning, NY, USA) supplemented with 10% FBS (Atlanta Biologicals, Minneapolis, MN, USA) and penicillin-streptomycin (Corning, NY, USA) at 5% CO₂/37 °C. BT-549 was maintained in RPMI-1640 medium supplemented with 10% FBS (Atlanta Biologicals, Minneapolis, MN, USA), 1% sodium pyruvate (Corning, NY, USA), 1% penicillin-streptomycin (Corning, NY, USA), 0.023 U/mL of Gibco™ Insulin, human recombinant zinc solution (Fisher Scientific, Hampton, NH, USA)). SK-BR-3 was maintained in Hyclone McCoy’s 5A Medium (GE Health care Life Sciences (Marlborough, MA, USA)) supplemented with 10% FBS (Atlanta Biologicals, Minneapolis, MN, USA) and 1% penicillin-streptomycin (Corning, NY, USA). 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), dimethyl sulfoxide (DMSO), crystal violet dye, and 16% paraformaldehyde (PFA) solution were purchased from Fisher Scientific (Hampton, NH, USA). All molecular biology kits and supplies were purchased from other commercial vendors which are listed at appropriate places throughout the manuscript.

Parvathaneni V., Chilamakuri R., Kulkarni N.S., Baig N.F., Agarwal S, & Gupta V. (2022). Exploring Amodiaquine’s Repurposing Potential in Breast Cancer Treatment—Assessment of In-Vitro Efficacy & Mechanism of Action. International Journal of Molecular Sciences, 23(19), 11455.

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Cell Culture Maintenance Protocol

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HEK-Blue IFN-α/β Cells, HEK 293, HEK 293T, and HeLa cells were maintained in DMEM (Cellgro) supplemented with 10% FBS (Atlanta Biologicals) and 1% penicillin-streptomycin (Gibco). U937 and NU-DUL-1 cells were maintained in RPMI (Cellgro) supplemented with 10% heat-inactivated FBS (Atlanta Biologicals) and 1% penicillin-streptomycin (Gibco). THP-1 cells were maintained in RPMI (Cellgro) supplemented with 10% heat-inactivated FBS (Atlanta Biologicals), 1% penicillin-streptomycin (Gibco), and 50 μM 2-mercaptoethanol (Sigma). All cell lines were maintained in a 5% CO₂ incubator at 37 °C.

Ritchie C., Cordova A.F., Hess G.T., Bassik M.C, & Li L. (2019). SLC19A1 is an importer of the immunotransmitter cGAMP. Molecular cell, 75(2), 372-381.e5.

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