Heat inactivated foetal bovine serum

Manufactured by Thermo Fisher Scientific

Sourced in United States, United Kingdom, Belgium

Heat-inactivated foetal bovine serum is a cell culture media supplement. It is derived from the blood of bovine fetuses and undergoes a heat-inactivation process to reduce the presence of contaminants.

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54 protocols using heat inactivated foetal bovine serum

Cell Culture Reagents and Protocols

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All cell and tissue culture media and supplements were purchased as sterile or were filter sterilised through a 0.20 µM filter. Heat-inactivated (hi) foetal bovine serum (FBS) and hi new born calf serum (NCS) were purchased from Gibco (Paisley, Scotland); hi horse serum (HS) from TCS Biosciences (Corby, England); penstrep (penicillin and streptomycin) and trypsin from Bio Whittaker (Wokingham, England); l-glutamine from BDH (Poole, England), gelatine from Sigma (St. Louis, USA). Glassware, distilled water (dH₂O) and phosphate buffered saline (PBS, Oxoid (Hampshire, England) were autoclaved prior to use. Sterile media were stored at 4 °C and used within 2 months. Plasticware were purchased as sterile from Greiner Bio-one, (Kremsmunster, Austria) unless otherwise stated. EPA purchased from IDS Ltd (Boldon, Tyne & Wear, UK) and Palmitate (Sigma-Aldrich, Poole, UK) were complexed with fatty-acid free bovine serum albumin (BSA) (Sigma-Aldrich, Poole, UK). Recombinant human TNF-α was purchased from Calbiochem (Nottingham, England); TRIZOL reagent (Invitrogen, Life Technologies), propidium iodide (PI) stain from BD Biosciences (San Diego, USA) and creatine kinase assay kits from Catachem (Bridgeport, USA). Recombinant human IGF-I was purchased from GroPep (Adelaide, Australia). BCA™ protein quantitation reagents were obtained from Pierce Chemicals (Chester, England, UK).

Saini A., Sharples A.P., Al-Shanti N, & Stewart C.E. (2016). Omega-3 fatty acid EPA improves regenerative capacity of mouse skeletal muscle cells exposed to saturated fat and inflammation. Biogerontology, 18(1), 109-129.

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Cell and Tissue Culture Supplies

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Al-Dabbagh S., McPhee J.S., Piasecki M., Stewart C.E, & Al-Shanti N. (2019). Soluble Factors Released From Activated T Lymphocytes Regulate C2C12 Myoblast Proliferation and Cellular Signaling, but Effects Are Blunted in the Elderly. The journals of gerontology. Series A, Biological sciences and medical sciences, 74(9).

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Cytotoxicity Assay with Cisplatin and Ethanol

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RPMI-1640 media, heat-inactivated foetal bovine serum (FBS), streptomycin, penicillin, cisplatin and ethanol were obtained from Thermo Fisher Scientific (Melbourne, Australia). Cell Counting Kit-8 (CCK-8) and TRIzol regent were purchased from Sigma-Aldrich (Sydney, Australia). TNFR1, NF-kB and Nrf2 primers were purchased from Bioneer Pacific (Melbourne, Australia), SensiFAST SYBR No-ROX qPCR and superscript III reverse transcriptase kits were purchased from Bioline (Sydney, Australia).

Nguyen D., Holien J., Dekiwadia C., Thrimawithana T., Piva T, & Huynh T. (2023). Momordica cochinchinensis (Gấc) Seed Extracts Induce Apoptosis and Necrosis in Melanoma Cells. Pharmaceuticals, 16(1), 100.

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Cell Culture Protocols for Retinal and Vero Cells

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Both the murine retinal photoreceptor cell line 661W and Vero cells were obtained from Osaka Bioscience Institute. Both 661W and Vero cells were grown in Dulbecco’s modified Eagle’s medium (DMEM)/Ham’s F12 with l-glutamine and phenol red (661W) (Fujifilm Wako Pure Chemical Corporation) or high-glucose DMEM with l-glutamine, phenol red, HEPES, and sodium pyruvate (Vero) (Fujifilm Wako Pure Chemical Corporation), supplemented with 10% heat-inactivated foetal bovine serum (FBS) (Thermo Fisher Scientific) in a humidified incubator with 5% CO₂. Retinal microvascular endothelial cells of BALB/c mice were obtained from Cell Biologics and were cultured in complete mouse endothelial cell medium/w Kit (Cell Biologics) supplemented with 10% heat inactivated FBS (Thermo Fisher Scientific) in a humidified incubator with 5% CO₂.

Shirahama S., Onoguchi-Mizutani R., Kawata K., Taniue K., Miki A., Kato A., Kawaguchi Y., Tanaka R., Kaburaki T., Kawashima H., Urade Y., Aihara M, & Akimitsu N. (2020). Long noncoding RNA U90926 is crucial for herpes simplex virus type 1 proliferation in murine retinal photoreceptor cells. Scientific Reports, 10, 19406.

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Culturing Human Umbilical Vein Endothelial and Rat Schwann Cells

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Human umbilical vein endothelial cells (HUVECs; PromoCell) were grown in complete Endothelial Cell Growth Media (EGM) (PromoCell). They were used between passages 4 and 10. Schwann cells were from the rat Schwann cell line SCL4.1/F7 (Health Protection Agency) and were grown in culture medium (Dulbecco's Modified Eagle's Medium , DMEM; Gibco). They were used between passages 4 and 20. All media were supplemented with 10% v/v heat-inactivated foetal bovine serum (FBS; Thermo Fisher Scientific) and 1% v/v Penicillin/Streptomycin (Gibco) and replaced every 2 days. All cell cultures and subsequent experiments were maintained in a humidified incubator with 5% CO 2 /95% air at 37 ºC.

Muangsanit P., Roberton V., Costa E, & Phillips J.B. (2021). Engineered aligned endothelial cell structures in tethered collagen hydrogels promote peripheral nerve regeneration. Acta biomaterialia, 126.

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High-throughput Screening for Candida Inhibitors

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High-throughput screening was performed at the Chemical Genetics Unit, Department of Microbiology, Research Center of Infection and Immunology, Li Ka Shing Faculty of Medicine, University of Hong Kong, on a library with 50,240 small molecules (ChemBridge, San Diego, CA, USA) to identify inhibitors of Y-H transition in C. albicans SC5314, as was previously described [29] (link). C. albicans SC5314 were seeded at 5×10³ cells per well in complete yeast peptone dextrose (YPD) (1% yeast extract, 2% peptone, 2% glucose/dextrose) supplemented with 20% heat-inactivated foetal bovine serum (Invitrogen, Carlsbad, CA, USA) in a total volume of 50 µl in 384-well microtitre plates. The small molecules were dissolved in dimethyl sulfoxide (DMSO) and were added to the wells at final concentration of 20 µg/ml, whereas the controls contained the same amount of DMSO but without small molecules. Assay plates were incubated at 37°C in 5% CO₂ for 12 h. Morphologies of the Candida were scored using a Leica DMIL inverted microscope equipped with DC300F digital imaging system (Leica Microsystems, Heidelberg, Germany). Small molecules with lower scores than that of the control (yeast-to-hypha transition inhibitors) were selected as primary hits.

Wong S.S., Kao R.Y., Yuen K.Y., Wang Y., Yang D., Samaranayake L.P, & Seneviratne C.J. (2014). In Vitro and In Vivo Activity of a Novel Antifungal Small Molecule against Candida Infections. PLoS ONE, 9(1), e85836.

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Culturing Human Umbilical Vein Endothelial Cells

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Human umbilical vascular endothelial cells (HUVECs) were purchased from the American Type Cell Culture Collection (ATCC, lot number: CRL-1730) and were cultured in RPMI-1640 medium (GIBCO, Invitrogen Inc., Carlsbad, CA, USA) containing 10% heat-inactivated foetal bovine serum (GIBCO, Invitrogen Inc.) and 2 mM L-glutamine at 37°C in a humidified atmosphere with 5% CO₂.

Zeng H., Kong X., Zhang H., Chen Y., Cai S., Luo H, & Chen P. (2020). Inhibiting DNA methylation alleviates cigarette smoke extract-induced dysregulation of Bcl-2 and endothelial apoptosis. Tobacco Induced Diseases, 18, 51.

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Leishmania Parasite Isolation and DNA Extraction

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Parasite samples were collected by wound aspiration from Leishmania-infected patients after one month of lesion appearance using 200–300 µl of sterile PBS. Wound aspirates were transferred to plastic flasks (in triplicate) containing Leishmania culture medium M199 (Gibco, Grand Island, NY, USA) supplemented with 15% heat-inactivated foetal bovine serum (Invitrogen), 1.5% BME vitamins (Sigma-Aldrich, Saint Louis, MO, USA) and 25 µg/ml gentamycin sulphate (Sigma-Aldrich, Saint Louis, MO, USA). Leishmania cultures were maintained for several days at 27 °C and parasite DNA was extracted from logarithmic phase cultures using the DNeasy Blood and Tissue kit (Qiagen, Germantown, MD, USA) following the manufacturer’s instructions.

Al-Salem W.S., Solórzano C., Weedall G.D., Dyer N.A., Kelly-Hope L., Casas-Sánchez A., Alraey Y., Alyamani E.J., Halliday A., Balghonaim S.M., Alsohibany K.S., Alzeyadi Z., Alzahrani M.H., Al-Shahrani A.M., Assiri A.M., Memish Z, & Acosta-Serrano Á. (2019). Old World cutaneous leishmaniasis treatment response varies depending on parasite species, geographical location and development of secondary infection. Parasites & Vectors, 12, 195.

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Oligo Library Screening in Cell Lines

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The 76nt random oligonucleotide library, 5'-ATC CAG AGT GAC GCA GCA-40nt-TGG ACA CGG TGG CTT AGT-3' FITC labelled upstream primer 5'FITC-ATC CAG AGT GAC GCA GCA, and Biotin labelled downstream primer 5'Biotin-ACT AAG CCA CCG TGT CCA-3' were synthesised by Huada Biotech (Beijing, China) Co., Ltd. U2-OS cells, HOS cells, and HT-1080 cells were purchased from Boshide Bioengineering Co., Ltd. McCoy's 5A medium and high-sugar DMEM medium were all purchased from Boshide Bioengineering Co., Ltd (Wuhan, China). Yeast tRNA (Fluka Analytical, USA), heat-inactivated foetal bovine serum (FBS Invitrogen, USA), Streptavidin Sepharose ® HP (GE healthcare, USA), RPMI-1640 medium (GIBCO), 2x Taq PCR Master Mix, 50 bp DNA Ladder (Tiangen, Japan), and an agarose gel recovery kit (Dingguo, China) were also used.

Wang H., Liang J., Ma Y., Zhou L., Zhang Y., & Li M. (2020). Development and Validation of a Novel Quality Control Methodology on Screening Out for Human Osteosarcoma Cell Aptamers.

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Screening of Oligonucleotide Libraries in Cell Lines

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The 76nt random oligonucleotide library, 5'-ATC CAG AGT GAC GCA GCA-40nt-TGG ACA CGG TGG CTT AGT-3',FITC labelled upstream primer 5'FITC-ATC CAG AGT GAC GCA GCA, and Biotin labelled downstream primer 5'Biotin-ACT AAG CCA CCG TGT CCA-3' were synthesised by Huada Biotech (Beijing, China) Co., Ltd. U2-OS cells, HOS cells, and HT-1080 cells were purchased from Boshide Bioengineering Co., Ltd. McCoy's 5A medium and high-sugar DMEM medium were all purchased from Boshide Bioengineering Co., Ltd (Wuhan, China). Yeast tRNA (Fluka Analytical, USA), heat-inactivated foetal bovine serum (FBS Invitrogen, USA), Streptavidin Sepharose® HP (GE healthcare, USA), RPMI-1640 medium (GIBCO), 2x Taq PCR Master Mix, 50 bp DNA Ladder (Tiangen, Japan), and an agarose gel recovery kit (Dingguo, China) were also used.

Wang H., Li M., Ma Y., Zhou L., Zhang Y., & Li M. (2021). Development and validation of a novel quality control methodology on screening out for human osteosarcoma cell aptamers.

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