Fetal bovine serum (fbs)

Manufactured by Sartorius

Sourced in Israel, United States, China, Japan, Germany, Australia, United Kingdom, Canada, France, Belgium

The FBS (Fetal Bovine Serum) is a cell culture media supplement used to promote the growth and proliferation of cells in vitro. It is a complex mixture of proteins, growth factors, and other components derived from the blood of fetal bovine. The FBS provides essential nutrients and growth factors that support the survival and expansion of various cell types in cell culture applications.

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2 887 protocols using fetal bovine serum (fbs)

Culturing Diverse Cell Lines for Research

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ARP1 and H929 human MM cell lines and the murine MM cell line, 5TMM3VT, were purchased from the Cell Resource Center of the Shanghai Institute of Biochemistry and Cell Biology at the Chinese Academy of Sciences. Cells were cultured in RPMI 1640 (Biological Industries, Beit Haemek, Israel), supplemented with 10% fetal bovine serum (FBS; Biological Industries, Israel) and 1% penicillin/streptomycin (P/S).
The pre-osteoblast murine cell line MC3T3-E1 was purchased from the Cell Resource Center of the Shanghai Institute of Biochemistry and Cell Biology at the Chinese Academy of Sciences, and cultured in alpha modified Eagle’s medium (α-MEM; Biological Industries, Israel) supplemented with 10% FBS (Biological Industries, Israel) and 1% (P/S). Cells were routinely subcultured using 0.05% trypsin/EDTA (Biological Industries, Israel) upon reaching 80–90% confluence.
The pre-osteoclast murine cell line Raw264.7 was purchased from the ATCC and cultured in Dulbecco’s modified Eagle medium (DMEM; Biological Industries, Israel), supplemented with 10% FBS (Biological Industries, Israel) and 1% P/S. After reaching 80–90% confluence, cells were routinely subcultured using 0.05% trypsin/EDTA (Biological Industries, Israel). All cells were maintained at 37°C in a humidified atmosphere of 5% CO2. Medium was changed every 2 days.

Hou J., Qian J., Li Z., Gong A., Zhong S., Qiao L., Qian S., Zhang Y., Dou R., Li R., Yang Y, & Gu C. (2020). Bioactive Compounds from Abelmoschus manihot L. Alleviate the Progression of Multiple Myeloma in Mouse Model and Improve Bone Marrow Microenvironment. OncoTargets and therapy, 13, 959-973.

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Culturing Human Bladder Cancer and Normal Urothelial Cells

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Human bladder cancer (5637 and 253 J), and normal human urothelial (SV-HUC-1) cell lines were purchased from American Type Culture Collection (ATCC, Rockville, Maryland, USA). The 5637 cell lines were cultured in a RPMI-1640 medium supplemented with 10% fetal bovine serum (FBS; Biological Industries, USA). The 253 J cell line was cultured in Dulbecco’s modified eagle’s medium (DMEM) supplemented with 10% FBS (Biological Industries, USA). The SV-HUC-1 cell line was cultured in Ham's F-12 K medium supplemented with 10% FBS (Biological Industries, USA). All cell lines were cultured at 37 ℃, aired with 5% CO2 and 95% humidity in a cell incubator.

Jiang Y., Zhang M., Wang L., Zhang L., Ma M., Jing M., Li J., Song R., Zhang Y., Yang Z., Zhang Y., Pu Y., Qu X, & Fan J. (2023). Potential mechanisms of osthole against bladder cancer cells based on network pharmacology, molecular docking, and experimental validation. BMC Complementary Medicine and Therapies, 23, 122.

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Culturing Diverse Human Cell Lines

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RL95-2, HEC-1A, and Ishikawa cell lines were purchased from the Food Industry Research and Development Institute of Taiwan (Hsinchu, Taiwan). RL95-2 cells were obtained from Caucasians, and the remaining cell lines were derived from Asians. The culture medium for RL95-2 consisted of 90% DMEM:F12 (1:1, Invitrogen, Thermo Fisher Scientific, Waltham, MA, USA) with 10 mM 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (Sigma–Aldrich), 5 μg/mL bovine insulin (Sigma–Aldrich), 2 g/mL sodium bicarbonate (Sigma–Aldrich), and 10% FBS (Biological Industries, Kibbutz, Israel). Ishikawa cells were cultured in minimal essential medium (MEM) with 2 mM glutamine, 1% nonessential amino acids (Sigma–Aldrich), and 5% FBS (Biological Industries). Finally, HEC-1A cells were grown in McCoy’s 5a medium modified (ATCC 30-2007, USA) with 10% FBS (Biological Industries).

Ding D.C., Chu T.Y, & Liu H.W. (2017). Reciprocal crosstalk between endometrial carcinoma and mesenchymal stem cells via transforming growth factor-β/transforming growth factor receptor and C–X–C motif chemokine ligand 12/C–X–C chemokine receptor type 4 aggravates malignant phenotypes. Oncotarget, 8(70), 115202-115214.

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Cell Line Characterization and Maintenance

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Wild‐type HCT116, MHCC‐97H, MCF‐7, HT29, and 293T cell lines were purchased from the Cell Bank of Chinese Academy of Sciences (Shanghai, China), and cultured in Dulbecco's modified Eagle's medium (Gibco, Life Technologies, Grand Island, NY) supplemented with 10% fetal bovine serum (FBS; Biological Industries, Beit Haemek, Israel) and 1% penicillin‐streptomycin. LoVo cell lines were purchased from the Cell Bank of Chinese Academy of Sciences (Shanghai, China), and cultured in F12K Ham's Kaighn's Modification medium (Macgene, Beijing, China), supplemented with 10% FBS (Biological Industries) and 1% penicillin‐streptomycin. p53‐null HCT116 (HCT116^p53null) cells were kindly provided by Dr. Bert Vogelstein at John Hopkins University School of Medicine, and maintained in McCoy's 5A medium (Gibco, Grand Islands, NY) with 10% FBS (Biological Industries) and 1% penicillin‐streptomycin. Cell lines were verified using short‐tandem repeat profiling method, and were tested periodically for mycoplasma contamination by using Mycoplasma Detection Kit‐QuickTest (Biotool, Houston, TX).

Li Y., Li J., Li Z., Wei M., Zhao H., Miyagishi M., Wu S, & Kasim V. (2022). Homeostasis Imbalance of YY2 and YY1 Promotes Tumor Growth by Manipulating Ferroptosis. Advanced Science, 9(13), 2104836.

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Cell Culture and Gene Silencing Protocol

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The KGN cell line (obtained from RIKEN BioResource Center, Ibaraki, Japan), a steroidogenic human granulosa-like tumor cell line^{59 (link)}, was cultured in DMEM/F12 (HyClone, Logan, UT, USA) supplemented with 10% fetal bovine serum (FBS; Biological Industries (BioInd), Beit Haemek, Israel) and 1% antibiotics (HyClone, Logan, UT, USA). The human embryonic kidney (HEK) 293T cell line and HeLa (human cervix carcinoma cell line) cell line were grown in DMEM High Glucose (HyClone, Logan, UT, USA) supplemented with 10% FBS (BioInd, Beit Haemek, Israel) and 1% antibiotics (HyClone, Logan, UT, USA). All cells were cultured at 37 ℃ in a humidified atmosphere of 5% CO₂ in air. MiR-379-5p mimics, specific siRNAs for PARP1/XRCC6 and negative control, were designed (siRNAs for PARP1 referred to Refs. ^{60 (link)} and ^{61 (link)}) and synthesized by Genepharma Inc (Shanghai, China). MiR-379-5p mimics and siRNA-MIX were transfected at 50 nM and 100 nM, respectively, using X-tremeGENE siRNA Transfection Reagent (La Roche Ltd, CH). Sequences were provided in Supplementary Table 2.

Dang Y., Wang X., Hao Y., Zhang X., Zhao S., Ma J., Qin Y, & Chen Z.J. (2018). MicroRNA-379-5p is associated with biochemical premature ovarian insufficiency through PARP1 and XRCC6. Cell Death & Disease, 9(2), 106.

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Isolation and Expansion of PDLSCs

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The extracted teeth were stored in Dulbecco’s modified Eagle’s medium (DMEM, HyClone, Logan, UT, USA) with 5% antibiotics (100 U/mL penicillin, 100 mg/mL streptomycin, Sigma Aldrich, St Louis, MO, USA) and quickly transported from the clinic to the laboratory. Then, single PDLSCs were acquired as previously described in our previous study (Du, Feng & Ge, 2016 (link)). Specifically, Primary PDLSCs were cultured with DMEM containing 20% fetal bovine serum (FBS, BioInd, Kibbutz, Israel) at 37 °C in a humidified atmosphere of 5% CO₂, and cells were trypsinized and passaged at a dilution ratio of 1:3 to expand the culture in 10% FBS medium upon the cell monolayer reached 80–90% confluence. Fourth passage cells were used in all experiments.

Kang W., Du L., Liang Q., Zhang R., Lv C, & Ge S. (2021). Transcriptome analysis reveals the mechanism of stromal cell-derived factor-1 and exendin-4 synergistically promoted periodontal ligament stem cells osteogenic differentiation. PeerJ, 9, e12091.

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Breast Cancer Cell Line Cultivation

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T47D, MCF7, BT549, and HS578T were procured from FuHeng Biology (FuHeng, China) and authenticated by STR (short tandem repeat) matching analysis. LCC2 was induced by MCF7 to form a stable cell line treated with low dose of tamoxifen for 6 months. MCF-7, LCC2, and HS578T cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM, BioInd, Israel) supplemented with 10% fetal bovine serum (FBS, BioInd, Israel), and 1% penicillin/streptomycin (P/S, BioInd, Israel). T47D and BT549 cells were cultured in RPMI-1640 (BioInd, Israel) supplemented with 10% FBS, and 1% P/S. All these cells were incubated at 37 °C in a humidified atmosphere with 5% CO₂.

Liu S., Liu B., Zhao Q., Shi J., Gu Y., Guo Y., Li Y., Liu Y., Cheng Y., Qiao Y, & Liu Y. (2021). Down-regulated FST expression is involved in the poor prognosis of triple-negative breast cancer. Cancer Cell International, 21, 267.

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Isolation and Cryopreservation of PBMCs

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Peripheral blood mononuclear cells (PBMCs) were isolated from heparinized venous blood by centrifugation over Ficoll-Paque (Lymphoprep; Alere Technologies AS, Oslo, Norway). The cells were resuspended in a freezing solution containing 10% dimethyl sulfoxide (DMSO) (Sigma, St. Louis, MO, USA) and 90% fetal bovine serum (FBS) (Biological Industries, Kibbutz Beit Hemek, Israel) and were frozen at −80 °C in an iso-propanol-jacketed closed container overnight. The cells were then stored in liquid nitrogen until use. A human embryonic kidney (HEK-293T) cell line (ATCC, Manassas, VA, USA) was maintained in DMEM (low glucose) medium supplemented with 10% FBS, 4 mM L-glutamine, 50 units/mL penicillin, and 50 µg/mL streptomycin (Biological Industries, Kibbutz Beit Hemek, Israel). The cells were cultured at 37 °C in a humidified atmosphere and 5% CO₂ in the air.

Golan M., Krivitsky A., Mausner-Fainberg K., Benhamou M., Vigiser I., Regev K., Kolb H, & Karni A. (2021). Increased Expression of Ephrins on Immune Cells of Patients with Relapsing Remitting Multiple Sclerosis Affects Oligodendrocyte Differentiation. International Journal of Molecular Sciences, 22(4), 2182.

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Culturing and Transfection of Cell Lines

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MCF-10A, MCF-7, MDA-MB-231 and HepG2 cells were maintained in Dulbecco's modified Eagle's medium (Gibco, Life Technologies, Grand Island, NY) with 10% fetal bovine serum (Biological Industries, Israel) and 1% penicillin-streptomycin. HCT116 and HCT116^p53-/- cells were kindly provided by Dr. Bert Vogelstein at The John Hopkins University Medicine School [32 (link)], and maintained in McCoy's 5A medium (Gibco) with 10% fetal bovine serum (Biological Industries) and 1% penicillin-streptomycin. All cells were cultured at 37^°C in a humidified incubator with 5% CO2. Cells were transfected with indicated vectors using Lipofectamine 2000 (Invitrogen Life Technologies) according to the manufacturer's protocol. For gene-silencing experiments, 24 h after transfection, transfected cells were selected by using 1 μg/mL puromycin. All cell lines have been routinely tested for mycoplasma contamination using Mycoplasma Detection Kit-Quick Test (Biotool, Houston, TX). No cell line was found positive for mycoplasma.

Kasim V., Xie Y.D., Wang H.M., Huang C., Yan X.S., Nian W.Q., Zheng X.D., Miyagishi M, & Wu S.R. (2017). Transcription factor Yin Yang 2 is a novel regulator of the p53/p21 axis. Oncotarget, 8(33), 54694-54707.

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HepG2 and LO2 Cell Culture Protocol

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The human hepatoma HepG2 cell line was obtained from American Type Culture Collection (Bethesda, MD, USA). Cells were grown in DMEM medium (Gibco, USA) supplemented with 10% fetal bovine serum (Biological Industries, Israel), 100 units/mL penicillin and 60 μg/mL streptomycin (Gibco, USA) at 37°C in a humidified atmosphere comprised of 95% air and 5% CO₂. Human normal liver cell LO2 was purchased from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China). Cells were grown in RPMI-1640 medium (Gibco, USA) supplemented with 10% fetal bovine serum (Biological Industries, Israel). Exponentially growing culture was maintained in a humidified atmosphere comprised of 95% air and 5% CO₂ at 37°C.
Specific pathogen-free male BALB/c nude mice, weighing 18–22 g, between 4 and 6 weeks of age, were purchased from Shanghai Slac Laboratory Animal Co. LTD (Shanghai, China). The mice were raised under controlled temperature (26-28°C, 55 ± 5% humidity) and daily light intensity (12 h of light), and were fed with standard laboratory food and water adlibitum. The animal experiments were conducted with the approval of the Animal Ethics Committee of China Pharmaceutical University.

Zhao P., Chen L., Li L.H., Wei Z.F., Tong B., Jia Y.G., Kong L.Y., Xia Y.F, & Dai Y. (2014). SC-III3, a novel scopoletin derivative, induces cytotoxicity in hepatocellular cancer cells through oxidative DNA damage and ataxia telangiectasia-mutated nuclear protein kinase activation. BMC Cancer, 14, 987.

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