Fetal bovine serum (fbs)

Manufactured by Dutscher

Sourced in France, United States, United Kingdom

Fetal bovine serum is a common cell culture media supplement. It is derived from the blood of bovine fetuses and provides a complex mixture of proteins, growth factors, and other components that support the growth and proliferation of cells in vitro.

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Lab products found in correlation

Streptomycin, by Thermo Fisher Scientific (10 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (9 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (8 mentions) Penicillin, by Thermo Fisher Scientific (7 mentions) Penicillin streptomycin, by Dutscher (5 mentions) Streptomycin, by Dutscher (4 mentions) Dulbecco phosphate buffered saline (dpbs), by Thermo Fisher Scientific (4 mentions) Penicillin streptomycin, by Lonza (4 mentions) Rpmi 1640, by Dutscher (4 mentions) L glutamine, by Thermo Fisher Scientific (4 mentions) L glutamine, by Merck Group (4 mentions) Zell shield, by Dutscher (3 mentions) Venorgem classic mycoplasma detection, by Minerva Biolabs (3 mentions) Penicillin, by Dutscher (3 mentions) High glucose dmem medium, by Dutscher (3 mentions) Zeocin, by InvivoGen (3 mentions) Dulbecco s modified eagle s medium, by Dutscher (3 mentions) Dulbecco modified eagle medium (dmem), by Lonza (3 mentions) Sw480, by American Type Culture Collection (3 mentions) Sw620, by American Type Culture Collection (3 mentions)

87 protocols using fetal bovine serum (fbs)

Cell Line Cultivation Protocols for CD20 and HER2 Targeting

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The Daudi cell line (ATCC Cat#CCL-213) and SK-BR-3 cell line (ATCC Cat#HTB-30) were used for experiments targeting CD20 and HER2, respectively. Daudi cells were cultured in RPMI-1640 (Sigma-Aldrich, St. Louis, MO, USA) containing 10% FBS (Dominique Dutscher, France), 100 U/mL penicillin and 100 µg/mL streptomycin (Sigma-Aldrich, St. Louis, MO, USA). SK-BR-3 cells were maintained in DMEM Glutamax culture medium (Thermo Scientific, Waltham, MA, USA) containing 10% FBS (Dominique Dutscher, France), 100 U/mL penicillin and 100 µg/mL streptomycin (Sigma-Aldrich, St. Louis, MO, USA).
MDCKII cells expressing human FcRn [15 (link)] were maintained in RPMI 1640 culture medium (Sigma-Aldrich, St. Louis, MO, USA) supplemented with 10% heat-inactivated FBS (Dominique Dutscher, Brumath, France), 2 mM l-glutamine (Sigma-Aldrich, St. Louis, MO, USA), 500 units of 0.5 mg/mL penicillin-streptomycin (Sigma-Aldrich, St. Louis, MO, USA) and 4 mg/mL G418 (Thermofisher Scientific, Waltham, MA, USA).

Aubrey N., Gouilleux-Gruart V., Dhommée C., Mariot J., Boursin F., Albrecht N., Bergua C., Croix C., Gilotin M., Haudebourg E., Horiot C., Matthias L., Mouline C., Lajoie L., Munos A., Ferry G., Viaud-Massuard M.C., Thibault G, & Velge-Roussel F. (2022). Anticalin N- or C-Terminal on a Monoclonal Antibody Affects Both Production and In Vitro Functionality. Antibodies, 11(3), 54.

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Cell Culture Protocols for Lung and Pharyngeal Cell Lines

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The cell lines used in this study were maintained in an atmosphere containing 5% CO₂ at 37 °C in the culture medium recommended by ATCC. Human epithelial lung A549 cells (ATCC, CCL-185) and murine macrophages J774A.1 (ATCC, TIB-67) were cultured in DMEM high glucose medium (Dominique Dutscher, Brumath, France) supplemented with 10% of heat-inactivated fetal bovine serum (Dominique Dutscher). Human pharyngeal epithelial FaDu cells (ATCC, HTB-43) were cultured in MEM medium (Dominique Dutscher) supplemented with 10% of heat-inactivated fetal bovine serum.

Guilhen C., Miquel S., Charbonnel N., Joseph L., Carrier G., Forestier C, & Balestrino D. (2019). Colonization and immune modulation properties of Klebsiella pneumoniae biofilm-dispersed cells. NPJ Biofilms and Microbiomes, 5, 25.

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Human AML Cell Lines and Primary Samples

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Human AML cell lines MOLM14, KG1, KG1a and OCI-AML3 were purchased from the Leibniz Institute DSMZ-German Collection of Microorganisms and Cell cultures (Braunschweig, Germany). OCI-AML3 and MOLM14 Cell lines were grown in RPMI-1640 medium with Glutamax supplemented with 10% fetal bovine serum (Dominique Dutscher, Brumath, France) while KG1 and KG1a cells were grown in Iscoves-modified Dulbecco medium (IMDM) containing 20% fetal bovine serum. Frozen samples from AML patients have been obtained after informed consent and stored at the HIMIP collection (BB18 0033-00060). According to the French law, HIMIP collections has been declared to the Ministry of Higher Education and Research (DC 2008-307 collection 1) and obtained a transfer agreement (AC 2008-129) after approbation by the “Comité de Protection des Personnes Sud-Ouest et Outremer II” (ethical committee). Peripheral blood or bone marrow samples were frozen in fetal calf serum with 10% DMSO and stored in liquid nitrogen. AML primary samples were cultured in IMDM containing 20% fetal bovine serum and 100 units/mL of penicillin and streptomycin. For some experiments, fresh leukemic blasts recovered at diagnosis were immediately treated with ethanol, idarubicin 10 nM, DDA 2.5 µM or both DDA and idarubicin for 48 h. Patient’s characteristics are shown in Figure S2A.

Mouchel P.L., Serhan N., Betous R., Farge T., Saland E., De Medina P., Hoffmann J.S., Sarry J.E., Poirot M., Silvente-Poirot S, & Récher C. (2020). Dendrogenin A Enhances Anti-Leukemic Effect of Anthracycline in Acute Myeloid Leukemia. Cancers, 12(10), 2933.

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Culture and Maintenance of Cancer Cell Lines

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MDA-MB-231 (human mammary adenocarcinoma), A549 (human lung adenocarcinoma), SK-HEP-1 (human hepatocellular carcinoma), DU145 (human prostate adenocarcinoma) and the two human colon adenocarcinomas HCT116 and SW480 cell lines were cultured with high-glucose Dulbecco's modified Eagle's medium provided by Dutscher (Dutscher, Brumath, France) supplemented with 10% fetal bovine serum (Dutscher). PANC-1 (human pancreatic carcinoma) cell line was cultured in RPMI 1640 (Roswell Park Memorial Institute medium) provided by Dutscher and supplemented with 10% fetal bovine serum. All these cell lines were grown in an incubator at 37 °C and 5% CO₂.

Morlé A., Garrido C, & Micheau O. (2015). Hyperthermia restores apoptosis induced by death receptors through aggregation-induced c-FLIP cytosolic depletion. Cell Death & Disease, 6(2), e1633-.

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Culturing HEK-293T and Huh7 cells

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Human embryonic kidney HEK-293T cells (ATCC, CRL-1573) were cultured in minimum essential media (MEM) supplemented with 5% heat-inactivated fetal bovine serum (FBS) and human hepatoma Huh7 cells were cultured in DMEM medium supplemented with 10% heat-inactivated fetal bovine serum (FBS, Dutscher, Brumath, France). Both cell lines were cultured with antibiotics (PAN Biotech Dutscher, Brumath, France) at 37 °C under a 5% CO₂ atmosphere. Mouse anti-DDDDK tag monoclonal antibody (FLAG Ab) and mouse anti-6x(His) monoclonal antibody was purchased from Abcam (Cambridge, UK). The mouse anti-flavivirus NS1 antibody [D/2/D6/B7] (NS1 mAb^D/2/D6/B7), which reacts with a conformational epitope present on DENV-2, was purchased from Abcam (Cambridge, UK). The rabbit dengue virus type 2 NS1 polyclonal antibody PA5-33207 (NS1 Ab^PA5-32207) raised against the residues NS1-117/301 was purchased from (Thermo Fisher Scientific, Les Ulis, France). The rabbit anti-β actin polyclonal antibody was purchased from ABclonal (Massachusetts, USA). Goat anti-mouse IgG secondary antibody and Alexa Fluor Plus 488, was purchased from Invitrogen (Thermo Fisher, Les Ulis, France). Anti-mouse and anti-rabbit IgG HRP-conjugated secondary antibodies were purchased from Abcam (Cambridge, UK). DAPI was purchased from Euromedex (Souffelweyersheim, France).

Ogire E., Diaz O., Vidalain P.O., Lotteau V., Desprès P, & Roche M. (2021). Instability of the NS1 Glycoprotein from La Reunion 2018 Dengue 2 Virus (Cosmopolitan-1 Genotype) in Huh7 Cells Is Due to Lysine Residues on Positions 272 and 324. International Journal of Molecular Sciences, 22(4), 1951.

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Cell Culture of MRL/N-1 Cells

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MRL/N-1 cells were obtained from Tetsuya Kodama, Tohoku University School of Medicine, Japan. They were routinely cultured in Roswell Park Memorial Institute medium (RPMI) 1640 buffered-medium supplemented with 10% (v/v) fetal bovine serum (Dutscher), 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES) and gentamycin (10 µg/ml; Lonza BioWhittaker). The absence of mycoplasma contamination was systematically ensured.

Wang F., Bonam S.R., Schall N., Kuhn L., Hammann P., Chaloin O., Madinier J.B., Briand J.P., Page N, & Muller S. (2018). Blocking nuclear export of HSPA8 after heat shock stress severely alters cell survival. Scientific Reports, 8, 16820.

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Standardized in vitro cell culture protocol

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The human embryonic kidney 293 cell line (HEK 293, ECACC 85120602) was provided by Sigma-Aldrich (Saint-Quentin Fallavier, France). The hepatoma cell line HepG2 was provided by ECACC (85011430). JEG3 cell line (ECACC 92120308) was provided by CERDIC (Sophia-Antipolis, France). Cells were grown in phenol red-free EMEM (Abcys, Paris, France) containing 2 mM glutamine, 1% nonessential amino acid, 100 U/mL of antibiotics (a mixture of penicillin, streptomycin, and fungizone) (Lonza, Saint Beauzire, France), 10 mg/mL of liquid kanamycin (Dominique Dutscher, Brumath, France), and 10% Fetal Bovine Serum (PAA, les Mureaux, France). JEG3 cells were supplemented with 1 mM sodium pyruvate. Cells were grown with this medium at 37°C (5% CO₂, 95% air) during 48 h to 80% confluence, then washed, and exposed 24 h with serum-free EMEM to the APs or the formulations. Before treatment, all the pesticides were solubilized in a 100% DMSO solution, then diluted in serum-free medium to reach 0.5% DMSO (which had been previously proven not to be cytotoxic for the cells), and adjusted to a similar pH. This model was validated [29 (link)] and cytotoxic effects were similar in presence of serum but delayed by 48 h.

Mesnage R., Defarge N., Spiroux de Vendômois J, & Séralini G.E. (2014). Major Pesticides Are More Toxic to Human Cells Than Their Declared Active Principles. BioMed Research International, 2014, 179691.

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Cytotoxicity Evaluation of 1,4-Dihydroquinolines

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Cell lines MDA-MB-231 from triple-negative breast cancer and HeLa from cervical cancer were obtained from the American Type Culture Collection (ATCC; Rockville, MD, USA). They were cultured at 37 °C with 5% CO₂ in DMEM middle (Lonza, Basel, Switzerland), supplemented with 10% fetal bovine serum (Dutscher, Brumath, France), 1% Zell Shield (Dutscher) and 1% non-essentials amino-acids (Lonza). Cancer cells were plated in 96-well plates (2 × 10³ cells/wells) in triplicate, incubated at 37 °C with 5% CO₂ for 24 h, and treated with 1,4-Dihydroquinolines for 72 h at different concentrations. Cell viability was determined using CellTiter 96^® AQueous One Solution Cell Proliferation Assay (MTS, Promega, Madison, WI, USA). 20 µL reagent was added in each well and cells were incubated for 2 h at 37 °C/5% CO₂. Absorbance was measured at 490 nm (SPECTROstar Nano, BMG LABTECH, Ortenberg, Germany). Calculations of IC50 were performed using GraphPad Prism V7.0 software (GraphPad Software, La Jolla, CA, USA).

Marchand G., Wambang N., Pellegrini S., Molinaro C., Martoriati A., Bousquet T., Markey A., Lescuyer-Rousseau A., Bodart J.F., Cailliau K., Pelinski L, & Marin M. (2020). Effects of Ferrocenyl 4-(Imino)-1,4-Dihydro-quinolines on Xenopus laevis Prophase I - Arrested Oocytes: Survival and Hormonal-Induced M-Phase Entry. International Journal of Molecular Sciences, 21(9), 3049.

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Human Ovarian and Lung Cell Cultivation

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Human ovarian surface epithelial cells (HOSE cells or OECs, as mentioned in our study) and human embryonic lung fibroblasts (MRC5 cells) were purchased from Innoprot (Derio, Spain) and RD-Biotech (Besançon, France), respectively. OECs were cultivated in ovarian epithelial cell medium (serum-free) supplemented with ovarian epithelial cell growth supplement (OEpiCGS) and penicillin/streptomycin solution (P60132, Innoprot). MRC5 cells were cultivated in Dulbecco’s modified Eagle medium (Sigma-Aldrich, Saint-Louis, MO) supplemented with 10% fetal bovine serum (Dutscher, Bernolsheim, France) and penicillin (100 U/mL)-streptomycin (100 μg/mL) (Life Technologies, Eugene, OR). Cells were cultured under standard conditions (37 °C, 5% CO₂, 95% humidity). To note that, HOSE cells or OECs were isolated from healthy human ovaries, as mentioned in the technical data sheet (P10982, Innoprot). Cultures were verified as mycoplasma-free as determined by monthly screenings (VenorGem classic mycoplasma detection, Minerva Biolabs).

El Baba R., Haidar Ahmad S., Monnien F., Mansar R., Bibeau F, & Herbein G. (2023). Polyploidy, EZH2 upregulation, and transformation in cytomegalovirus-infected human ovarian epithelial cells. Oncogene, 42(41), 3047-3061.

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Cytotoxicity Screening of Compounds in HepG2 Cells

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HepG2 cells were purchased from American Type Culture Collection. The cells were cultured in Eagle’s Minimum Essential Medium (Ozyme, France) supplemented with 10% fetal bovine serum, 1X non-essential amino acids, 100 units/mL penicillin and 10 mg/mL streptomycin (Dutscher, Brumath, France). Cultures were kept under a CO₂/air (5%/95%) humidified atmosphere at 37 °C. Prior to the experiment, cells were seeded in 96-well culture plates at a density of 0.1 × 10⁶ cells per well. After 24 h of incubation, the culture medium was refreshed and 100 µL of the test compounds or DMSO (0.1%) were added. Compounds were tested at 4 concentrations (1–30 µM) in triplicate. For the MTT assay [25 (link)], after 24 h of treatment, cells were incubated with 50 µL MTT (0.5 mg/mL, Sigma Aldrich, city, France) at 37 °C for 2 h. Plates were centrifuged, MTT was removed and 100 µL DMSO was distributed per well. The absorbance at 570 nm was measured using microplate reader (brand). Cell viability was expressed as percentage of cell viability compared to controls (DMSO, 0.1%).

Dgachi Y., Bautista-Aguilera O.M., Benchekroun M., Martin H., Bonet A., Knez D., Godyń J., Malawska B., Gobec S., Chioua M., Janockova J., Soukup O., Chabchoub F., Marco-Contelles J, & Ismaili L. (2016). Synthesis and Biological Evaluation of Benzochromenopyrimidinones as Cholinesterase Inhibitors and Potent Antioxidant, Non-Hepatotoxic Agents for Alzheimer’s Disease. Molecules, 21(5), 634.

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