Foetal bovine serum (fbs)

Manufactured by Gemini Bio

Sourced in United States

Foetal bovine serum is a cell culture supplement derived from the blood of bovine fetuses. It provides a rich source of proteins, growth factors, and other nutrients essential for cell growth and proliferation in in vitro cell culture applications.

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12 protocols using foetal bovine serum (fbs)

Glutamine Deprivation Cell Model

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Glutamine deprivation cell model was established follow the method below, remove normal DMEM, and wash normal NSCLC cells or transfect cells with phosphate buffer saline (PBS). Adding DMEM (ThermoFisher Science, A14431) which without glutamine. In addition, 10% foetal bovine serum (Gemini Bio Products, Sacramento, California, USA) was added to the medium. The cells were cultured in this medium for 24 hours and continued to be used.

Xiao S., Nai‐dong W., Jin‐Xiang Y., Long T., Xiu‐Rong L., Hong G., Jie‐Cheng Y, & Fei Z. (2022). ANGPTL4 regulate glutamine metabolism and fatty acid oxidation in nonsmall cell lung cancer cells. Journal of Cellular and Molecular Medicine, 26(7), 1876-1885.

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Neuro-2a Cell Culture Protocol

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Mouse neuroblastoma Neuro-2a cells (a gift from Prof Wedong Le, Shanghai University of Medicine and Health Sciences, passage number 5 since received)were cultured in Dulbecco’s modified Eagle’s medium (HyClone, USA) containing 10% foetal bovine serum (Gemini Bio-Products, Canada) and 1% penicillin streptomycin (HyClone, USA), under a humidified atmosphere at 37 °C in an incubator containing 5% CO₂.

Song Y., Du Y., Zou W., Luo Y., Zhang X, & Fu J. (2018). Involvement of impaired autophagy and mitophagy in Neuro-2a cell damage under hypoxic and/or high-glucose conditions. Scientific Reports, 8, 3301.

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Culturing and Tracking BJ Fibroblasts

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Human BJ fibroblasts were provided by the Dillin laboratory (UC Berkeley) and validated by short tandem repeat identification (UC Berkeley Cell Culture Facility). These cells were mycoplasma-negative and cultured using DMEM + GlutaMAX media (ThermoFisher Scientific) supplemented with 1% penicillin/streptomycin (Life Technologies), 1× MEM non-essential amino acids (Life Technologies), and 10% foetal bovine serum (Gemini Bio-products). Cells were cultured at 37 °C with 5% CO₂ in a humidified incubator. All cells were between passage number 8 and 13.
For cell patterning, cells were detached using 0.25% trypsin-EDTA (Gemini Bio-products), re-suspended, and applied to the fibronectin-patterned gel slide and incubated at 37 °C for 1 h to allow for cell attachment. The gels were then rinsed with PBS to remove unadhered cells and cultured in culturing medium overnight (to allow for recovery of receptor proteins) or up to four days (to obtain 2000 cell per cm² density). For studies comparing GFP expression, BJ fibroblasts were first transduced with GFP using a BacMam GFP Transduction kit (Thermo Fischer Scientific) at a 5% (v/v) concentration. Cells were then used 24 h after transduction.
For fluorescence-labelling of cells, BJ fibroblasts were stained using Cytopainter Cell Tracking Staining Kit (ABCAM) according to the manufacturer’s instructions.

Su E.J, & Herr A.E. (2017). Electrophoretic cytometry of adherent cells. Lab on a chip, 17(24), 4312-4323.

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Cell Culture Media Preparation

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D10 medium was prepared by adding 50 ml heat-inactivated foetal bovine serum (Gemini Bio-Products, West Sacramento, CA, USA, Cat. No. 900-208) and 5.5 ml 100X L-Glutamine:Penicillin:Streptomycin solution (Gemini Bio-Products Cat. No. 400-110) to 500 ml Dulbecco's modified Eagle's medium without L-glutamine (Mediatech, Herndon, VA, USA, Cat. No. 15-013-CV). R10 medium was prepared by adding the same two components to 500 ml RPMI 1640 medium without L-glutamine (Mediatech Cat. No. 15-040). HEK293T cells, hereafter ‘293T,’ (ATCC, Manassas, VA, USA, Cat. No. CRL-1268) were maintained in D10 medium, and K562 (ATCC Cat. No. CCL-243) cells were maintained in R10 medium.

Cooper A.R., Lill G.R., Gschweng E.H, & Kohn D.B. (2014). Rescue of splicing-mediated intron loss maximizes expression in lentiviral vectors containing the human ubiquitin C promoter. Nucleic Acids Research, 43(1), 682-690.

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Characterization of Lung Cell Lines

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The human normal lung epithelial cell line, BEAS-2B (cat. no. HTX2075), was obtained from Otwo Biotech (Shenzhen Haodihuatuo Biotechnology Co., Ltd.). The lung cancer cell line, A549 (cat. no. CCL-185), and the 293T cell line (cat. no. CRL-11268) were obtained from the American Type Culture Collection. The lung cancer cell line, H460 (cat. no. CL-0299), was obtained from Procell Life Science & Technology Co., Ltd. All cell lines were authenticated by their suppliers through STR profiling. The A549, BEAS-2B, and 293T cells were cultured in DMEM (Gibco; Thermo Fisher Scientific, Inc.) with 10% foetal bovine serum (Gemini Bio Products), 100 U/ml penicillin and 100 μg/ml streptomycin at 37°C with 5% CO₂. H460 cells were cultured in RPMI-1640 medium (Gibco; Thermo Fisher Scientific, Inc.) with the same supplements and conditions.

Tai F., Zhai R., Ding K., Zhang Y., Yang H., Li H., Wang Q., Cao Z., Ge C., Fu H., Xiao F, & Zheng X. (2024). Long non-coding RNA lung cancer-associated transcript 1 regulates ferroptosis via microRNA-34a-5p-mediated GTP cyclohydrolase 1 downregulation in lung cancer cells. International Journal of Oncology, 64(6), 64.

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Caco-2 Barrier Dysfunction Assay

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Caco-2 cells were cultured in Eagle's Minimum Essential Medium supplemented with 10% heat-inactivated foetal bovine serum (Gemini Bioproducts) and 1% non-essential amino acids (Invitrogen). Cells were plated on transwell filters with a pore size of 0.4 μm (Corning, USA). TNF-α (20 ng/ml) and IFN-γ (10 ng/ml) were added basolaterally to monolayers with or without 100 nM FICZ.

Yu M., Wang Q., Ma Y., Li L., Yu K., Zhang Z., Chen G., Li X., Xiao W., Xu P, & Yang H. (2018). Aryl Hydrocarbon Receptor Activation Modulates Intestinal Epithelial Barrier Function by Maintaining Tight Junction Integrity. International Journal of Biological Sciences, 14(1), 69-77.

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Purification of Fetal Liver Erythroid Progenitors

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Purification of mouse foetal liver erythroid progenitors (TER119-negative cells) was performed as described previously29 (link)30 (link). In brief, foetal liver cells were isolated from C57BL/6 mice on embryonic day 13.5 and mechanically dissociated by pipetting in phosphate-buffered saline (PBS) containing 10% foetal bovine serum (Gemini Bio-Products). Single-cell suspensions were prepared by passing the dissociated cells through 40-μm cell strainers (BD Biosciences). Foetal liver cells were labelled with biotin-conjugated anti-TER119 antibody (1:100; eBioscience) and purified using the EasySep column-free cell isolation system (Stem Cell Technologies) according to the manufacturer’s instructions. Purified TER119-negative cells were cultured in erythropoietin (2 unit/ml, PROCRIT®) containing medium.

Zhao B., Tan T.L., Mei Y., Yang J., Yu Y., Verma A., Liang Y., Gao J, & Ji P. (2016). H2AX deficiency is associated with erythroid dysplasia and compromised haematopoietic stem cell function. Scientific Reports, 6, 19589.

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Muscle Tissue Regeneration Protocols

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Sample collection for CTX injection–alone group was performed as follows. The calf muscles of both legs of C57BL/6 J mice were injected with CTX as described earlier. At Day 3, Day 7 and Day 14 time points, mice were sacrificed and calf muscles from left and right sides were immediately harvested and snap-frozen in liquid nitrogen for later muscle tissue RNA and protein isolation, respectively. Uninjected mice served as Day 0 control (N = 3 for each time point).
A separate group of mice were used for muscle tissue–derived conditioned medium (CM) collection. The calf muscle of the right leg of C57 BL/6 J mice was injected with CTX as described earlier. On Day 2, mice were sacrificed and the calf muscles of both legs were processed as follows. Briefly, pooled muscle tissue samples (1 g) from either the control left leg or the CTX-injured right leg were cut into 3-mm pieces and cultured in 10 mL of growth medium [GM, high-glucose Dulbecco's Modiﬁed Eagle's Medium (Gibco, Gaithersburg, MD, USA) supplemented with 20% foetal bovine serum (Gemini Bio Products, West Sacramento, CA, USA) and 1% penicillin/streptomycin (Gibco)]. CM was changed and collected every day for three days and stored at −80 °C for later use.

Li L., Jiang Y., Lin H., Shen H., Sohn J., Alexander P.G, & Tuan R.S. (2019). Muscle injury promotes heterotopic ossification by stimulating local bone morphogenetic protein-7 production. Journal of Orthopaedic Translation, 18, 142-153.

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Fbxo4 Transgenic Mouse Generation

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Mouse breeding, genotyping, handling and treatment were carried out in accordance with IACUC protocols and University Laboratory Animal Research guidelines at the Medical University of South Carolina. Fbxo4 + / + , + /− and −/− transgenic mice in C57BL/6 background were developed by Vega Biolab (Philadelphia, PA). Six-week old male and female mice were utilised for breeding. At menstrual age day-14, mouse embryos were dissected out. The head and visceral organs and tissues were removed. Cells were maintained in MEFs medium on a 3T9 passaging protocol. MEFs medium contains Dulbecco modified Eagle’s medium (DMEM) with 10% foetal bovine serum (FBS) (Gemini Bio-Products), 2 mM glutamine, 0.1 mM nonessential amino acids, 55 μM β-mercaptoethanol, and 10 μg of gentamicin/ml. The genotyping primers were as follows: 1loxP forward, 5′-GGCAGAGCTTGAGTTTGCAACATTTCAGGTG-3′, and 3loxP reverse, 5′-TCCTGATCTTTGGAAATTCTTCCTCTGAGT-3′.

Qie S., Majumder M., Mackiewicz K., Howley B.V., Peterson Y.K., Howe P.H., Palanisamy V, & Diehl J.A. (2017). Fbxo4-mediated degradation of Fxr1 suppresses tumorigenesis in head and neck squamous cell carcinoma. Nature Communications, 8, 1534.

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Fluorescent Dye Labeling Protocol

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The N-hydroxysuccinimidyl (NHS) esters of different fluorescent dyes were purchased as follows: BDP, Cy3.5, Cy5 and Cy7.5 from Lumiprobe Corporation; ICG-Sulfo-OSu from AAT Bioquest Inc; BDP650 NHS ester from Life Technologies. Ultracentrifugal filter units (MWCO = 100 kDa) were obtained from Merck Millipore. Foetal bovine serum (FBS) was purchased from Gemini Bio-products Inc. Cell culture media, penicillin-streptomycin and 0.25% trypsin-EDTA were obtained from M&C gene technology Ltd. Paclitaxel (PTX) was purchased from Dalian Meilun Biotechnology Co. Ltd. Monomers, such as 2-(diisopropylamino) ethyl methacrylate (DPA-MA) and 2-aminoethyl methacrylate (AMA) were purchased from Polysciences Company. Initiator V65 was purchased from Yuanye Biotechnology Co. Ltd. Other reagents and solvents were from Thermo Fisher Scientific or Sigma-Aldrich Inc.

Yin Q., Pan A., Chen B., Wang Z., Tang M., Yan Y., Wang Y., Xia H., Chen W., Du H., Chen M., Fu C., Wang Y., Yuan X., Lu Z., Zhang Q, & Wang Y. (2021). Quantitative imaging of intracellular nanoparticle exposure enables prediction of nanotherapeutic efficacy. Nature Communications, 12, 2385.

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