The T24 cells were grown in McCoy’s 5A (Biological Industries) supplemented with 10% FBS (Capricorn Scientific). The cells were passaged twice per week, and the culture medium was changed with the same frequency. The LNCaP cells were maintained in RPMI media (Biological Industries) supplemented with 10% FBS (Capricorn Scientific) and 10% of its own conditioned medium (complete media). The cells were passaged once per week, and the medium was changed with the same frequency. The HaCaT cells (immortalized human keratinocytes) were cultured in Dulbecco’s modified Eagle medium (Biological Industries) supplemented with 10% FBS (Capricorn Scientific). The cells were passaged twice per week, and the medium was changed with the same frequency. The cell cultures were maintained under a humidified 5% CO2 atmosphere at 37 °C.
Fetal bovine serum (fbs)
Manufactured by Capricorn
Sourced in Germany, United States, United Kingdom, Belgium, Austria, Japan, Switzerland, India
FBS is a cell culture supplement derived from the blood serum of fetal bovine. It provides a rich source of proteins, growth factors, and other essential nutrients to support the growth and proliferation of a wide range of cell types in vitro.
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Lab products found in correlation
Penicillin streptomycin, by Thermo Fisher Scientific (32 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (31 mentions)
Penicillin streptomycin, by Capricorn (25 mentions)
Streptomycin, by Thermo Fisher Scientific (24 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (23 mentions)
Penicillin, by Thermo Fisher Scientific (21 mentions)
Dulbecco modified eagle medium (dmem), by Capricorn (20 mentions)
Streptomycin, by Capricorn (15 mentions)
Penicillin, by Capricorn (15 mentions)
L glutamine, by Thermo Fisher Scientific (12 mentions)
Penicillin streptomycin, by Merck Group (12 mentions)
Streptomycin, by Merck Group (11 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (11 mentions)
Penicillin, by Merck Group (10 mentions)
Dulbecco modified eagle medium (dmem), by Merck Group (10 mentions)
Rpmi 1640, by Thermo Fisher Scientific (10 mentions)
Trypsin edta, by Thermo Fisher Scientific (8 mentions)
L glutamine, by Capricorn (8 mentions)
Mcf 7, by American Type Culture Collection (8 mentions)
Antibiotic antimycotic solution, by Capricorn (7 mentions)
285 protocols using fetal bovine serum (fbs)
1
Cell Culture Maintenance Protocols
2
Cancer and Kidney Cell Culture Protocols
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All the four cancer cell lines (MCF-7: human breast adenocarcinoma cells; HeLa: human cervix adenocarcinoma cells; Caco-2: human epithelial colorectal adenocarcinoma cells; A549: human epithelial lung adenocarcinoma cells) were obtained from the American Type Culture Collection (ATCC) (Rockville, MD, USA). These cells were grown at 37 °C with 5% CO2 in a humidified environment in Dulbecco’s Modified Eagle’s Medium (DMEM) high glucose (4.5 g/L) containing L-glutamine (4 mM) and sodium-pyruvate (Hyclone™) supplemented with 10% (v/v) fetal bovine serum (Capricorn Scientific Gmbh, South America). African green monkey (Vero) kidney cell lines (also obtained from ATCC) were maintained at 37 °C and 5% CO2 in a humidified environment in DMEM high glucose (4.5 g/L) containing L-glutamine (Lonza, Belgium) and supplemented with 5% fetal bovine serum (Capricorn Scientific Gmbh, South America) and 1% gentamicin (Virbac, RSA). The RAW 264.7 murine macrophage cells (obtained from ATCC) were maintained in DMEM high glucose (4.5 g/L) containing L-glutamine (Lonza, Belgium) and supplemented with 10% fetal bovine serum (Capricorn Scientific Gmbh, South America) and 1% penicillin/streptomycin/fungizone (PSF) solution under 5% CO2 humidified environment at 37 °C.
3
Culturing Cancerous and Non-Cancerous Cell Lines
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Cancerous cell lines (MCF-7: human breast adenocarcinoma cells; HepG2: human hepatocellular carcinoma cells; Caco-2: human epithelial colorectal adenocarcinoma cells; A549: human epithelial lung adenocarcinoma cells), obtained from the American Type Culture Collection (ATCC) (Rockville, MD, United States of America), were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM) high glucose (4.5 g/L) containing L-glutamine (4 mM) and sodium-pyruvate (Hyclone™) supplemented with 10% (v/v) fetal bovine serum (FBS) (Capricorn Scientific GmbH, South America), and incubated at 37°C with 5% CO2 in a humidified environment. African green monkey (Vero) kidney cells (also obtained from ATCC), a non-cancerous cell line, were grown in DMEM high glucose (4.5 g/L) containing L-glutamine (Lonza, Belgium) and supplemented with 5% FBS (Capricorn Scientific GmbH, South America) and 1% gentamicin (Virbac, RSA) in the same environment as cancer cells. The RAW 264.7 murine macrophage cells (obtained from ATCC) were cultured in DMEM high glucose (4.5 g/L) containing L-glutamine (Lonza, Belgium), supplemented with 10% FBS (Capricorn Scientific GmbH, South America) and 1% penicillin/streptomycin/fungizone (PSF) solution, and kept at 37°C in a 5% CO2 humidified environment.
4
Culturing Breast Cancer Cell Lines
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The human breast cancer cell lines AMJ13 [20] , MCF7, MDAMB, and CAL51, as well as mouse embryo fibroblasts (MEF) were supplied by Cell Bank Unit. AMJ13 cells were cultured in RPMI-1640 medium (USbiological, USA) supplemented with 10% fetal bovine serum (FBS) (Capricorn Scientific, Germany), 100 units/mL penicillin, and 100 μg/ mL streptomycin. MCF7, MDAB, and CAL51 cells were cultured in Minimum Essential medium (MEM) (USbiological) supplemented with 10% FBS (Capricorn-Scientific, Germany), 100 μg/mL streptomycin, and 100 units/mL penicillin. The cells were incubated at 37 °C in a humidified environment and 5% CO 2 .
5
Cell Culture and Protein Expression
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The non-small cell lung cancer cell line H1299 was cultured in RPMI medium 1640 (Gibco), containing 10% FBS (Capricorn Scientific), 100 U/ml penicillin (Gibco) and 100 µg/ml streptomycin (Gibco) at 37 °C and 5% CO2. H1299 cell line was obtained from ATCC. T-REx-HeLa cell line was cultured in DMEM medium (Gibco), containing 10% FBS (Capricorn Scientific), 4 µg/ml blasticidin (Gibco), 333 µg/ml Zeocin (Gibco), 100 U/ml penicillin (Gibco), 100 µg/ml streptomycin (Gibco) and 1 mM pyruvate (Gibco) at 37 °C and 5% CO2. The T-REx-HeLa cell line was a gift from Christian Behrends (Munich Cluster for Systems Neurology (SyNergy), Ludwig-Maximilians-University (LMU), Munich, Germany).
All cell lines used in this study were routinely tested for mycoplasma contaminations.
For recombinant protein expression, H1299 cells in medium without antibiotics were transfected using Lipofectamine 2000 transfection reagent (Thermo Fisher Scientific) according to the manufacturer’s recommendation. 6 h after transfection the medium was exchanged to standard H1299 culturing medium.
All cell lines used in this study were routinely tested for mycoplasma contaminations.
For recombinant protein expression, H1299 cells in medium without antibiotics were transfected using Lipofectamine 2000 transfection reagent (Thermo Fisher Scientific) according to the manufacturer’s recommendation. 6 h after transfection the medium was exchanged to standard H1299 culturing medium.
6
Cell Culture Protocol for Astrocytoma and PC-12 Cells
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Astrocytoma (1321N1) cells (obtained from Sigma-Aldrich, acc. no. 86030402) were cultured in Gibco DMEM medium (Fisher Scientific, Inc., Waltham, MA, USA) containing 10% heat-inactivated FBS (Capricorn) [11 (link)]. Rat pheochromocytoma cells (PC-12, adherent variant) purchased from the European Collection of Authenticated Cell Cultures (ECACC) general collection were grown in Gibco RPMI-1640 (Fisher Scientific, Hampton, NH, USA) medium containing 10% horse serum (Capricorn Scientific GmbH, Ebsdorfergrund, Germany) and 5% heat-inactivated fetal bovine serum-FBS (Capricorn). The media were supplemented with penicillin (0.15 mM), streptomycin (86 µM), and glutamine (2 mM) [23 (link)]. The cells were incubated at 37 °C in a humidified environment of 7.5% CO2 and were routinely passaged every 3–4 days. Collagen type IV (Sigma C5533) was coated on 96-well plates and left for 6 h or more before using the plates whenever seeding PC-12 cells.
7
Vero E6 Cell Culture and SARS-CoV-2 Variant Propagation
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Vero E6 (ATCC CRL-1586) cells were maintained in Dulbecco’s modified Eagle’s medium (DMEM, HyClone, Cytiva, USA), supplemented with 10% or 2% of heat-inactivated fetal bovine serum (FBS, Capricorn Scientific, Germany), L-glutamine (4 mM) and penicillin/streptomycin solution (100 IU/mL; 100 μg/mL) (PanEco, Moscow, Russia).
The SARS-CoV-2 variants B.1.1.1 or PMVL-1 (GISAID EPI_ISL_421275) and Omicron BA.5 (hCoV-19/Russia/SPE-RII-25357S/2022) initially isolated from a nasopharyngeal swab were obtained from the State Collection of Viruses of the Gamaleya Center in Moscow and used in both challenge and Microneutralization Assay. Isolation and further propagation were performed in Vero E6 cells in DMEM (HyClone Cytiva, Austria) with 2% heat-inactivated FBS (Capricorn Scientific GmbH, Germany): the cells were infected at multiplicity of infection (MOI) = 0.01 and incubated at 37°C in 5% CO2. The culture medium was collected at 72 h and clarified by centrifugation at 9000 g for 10 min at +4°C. The culture medium containing the virus was aliquoted, frozen and stored at −80°C.
The SARS-CoV-2 variants B.1.1.1 or PMVL-1 (GISAID EPI_ISL_421275) and Omicron BA.5 (hCoV-19/Russia/SPE-RII-25357S/2022) initially isolated from a nasopharyngeal swab were obtained from the State Collection of Viruses of the Gamaleya Center in Moscow and used in both challenge and Microneutralization Assay. Isolation and further propagation were performed in Vero E6 cells in DMEM (HyClone Cytiva, Austria) with 2% heat-inactivated FBS (Capricorn Scientific GmbH, Germany): the cells were infected at multiplicity of infection (MOI) = 0.01 and incubated at 37°C in 5% CO2. The culture medium was collected at 72 h and clarified by centrifugation at 9000 g for 10 min at +4°C. The culture medium containing the virus was aliquoted, frozen and stored at −80°C.
8
Characterization of HCMV Replication Assays
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Primary human foreskin fibroblasts (HFFs, derived from clinical samples, Children’s Hospital, Erlangen, Germany) were grown in Eagle’s minimal essential medium (MEM) supplemented with 1 × GlutaMAXTM (both Thermo Fisher Scientific, Waltham, MA, USA), 10 μg/mL gentamicin and 10% fetal bovine serum (FBS, Capricorn, Ebsdorfergrund, Germany). Human embryonic kidney epithelial cells (293Ts, ATCC, Manassas, VA, USA) were grown in Dulbecco’s modified Eagle medium (DMEM) supplemented with 1 × GlutaMAXTM (both Thermo Fisher Scientific, Waltham, MA, USA), 10 μg/mL gentamicin and 10% FBS (Capricorn, Ebsdorfergrund, Germany). Cultured cells were maintained at 37 °C, 5% CO2 and 80% humidity. All cell culture was regularly monitored for absence of mycoplasma contamination (Lonza™ Mycoalert™, Thermo Fisher Scientific, Waltham, MA, USA). Recombinant HCMV AD169 expressing green fluorescent protein (AD169-GFP, [61 (link)]) and recombinant ORF-UL97-deleted HCMV AD169 expressing green fluorescent protein (ΔUL97-GFP BAC213 [48 (link)]) were used for in vitro replication assays.
9
Propagation and Characterization of SARS-CoV-2
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Vero E6 (ATCC CRL-1586) cells were maintained in Dulbecco’s modified Eagle’s medium (DMEM, HyClone, Cytiva, USA), supplemented with 10% or 2% of heat-inactivated fetal bovine serum (FBS, Capricorn Scientific, Germany), L-glutamine (4 mM) and penicillin/streptomycin solution (100 IU/mL; 100 μg/mL) (PanEco, Moscow, Russia).
Cells from mouse spleen and lungs as well as marmoset PBMCs were seeded in complete Roswell Park Memorial Institute (RPMI) medium supplemented with 10% heat-inactivated FBS (Gibco, USA).
The SARS-CoV-2 strain B.1.1.1 or PMVL-1 (GISAID EPI_ISL_421275) and B.1.617.2 (Delta) initially isolated from a nasopharyngeal swab was obtained from the State Collection of Viruses of the Gamaleya Center in Moscow and used in both challenge and titration of NtAbs studies. Isolation and further propagation were performed in Vero E6 cells in DMEM (HyClone Cytiva, Austria) with 2% heat-inactivated FBS (Capricorn Scientific GmbH, Germany): the cells were infected at multiplicity of infection (MOI) = 0.01 and incubated at 37°C in 5% CO2. The culture medium was collected at 72 h and clarified by centrifugation at 9000 g for 10 min at +4°C. The culture medium containing the virus was aliquoted, frozen and stored at −80°C.
Cells from mouse spleen and lungs as well as marmoset PBMCs were seeded in complete Roswell Park Memorial Institute (RPMI) medium supplemented with 10% heat-inactivated FBS (Gibco, USA).
The SARS-CoV-2 strain B.1.1.1 or PMVL-1 (GISAID EPI_ISL_421275) and B.1.617.2 (Delta) initially isolated from a nasopharyngeal swab was obtained from the State Collection of Viruses of the Gamaleya Center in Moscow and used in both challenge and titration of NtAbs studies. Isolation and further propagation were performed in Vero E6 cells in DMEM (HyClone Cytiva, Austria) with 2% heat-inactivated FBS (Capricorn Scientific GmbH, Germany): the cells were infected at multiplicity of infection (MOI) = 0.01 and incubated at 37°C in 5% CO2. The culture medium was collected at 72 h and clarified by centrifugation at 9000 g for 10 min at +4°C. The culture medium containing the virus was aliquoted, frozen and stored at −80°C.
10
Cell Culture Protocol for Mammalian Infection Studies
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Mammalian cells used for infection experiments were BHK-21 (hamster kidney cells; CCVL L 0179) and A549 (human lung epithelial cells; ATCC® CCL-185™). Cells were cultured in Dulbecco’s Modified Eagle Medium (DMEM; Thermo Scientific Inc., Waltham, MA, USA) (A549) or Minimum Essential Medium (MEM; Capricorn Scientific, Ebsdorfergrund, Germany) (BHK-21), both supplemented with (1%) penicillin–streptomycin (100 units/mL) (PAN-Biotech GmbH, Aidenbach, Germany), (1%) L-glutamine (200 mM) (PAN-Biotech GmbH, Aidenbach, Germany), and 10% FBS (A549) or 5% FBS (BHK-21) (Capricorn Scientific, Ebsdorfergrund, Germany) at 37 °C and 5% CO2. Cell cultures were split twice weekly to prevent cells from reaching full confluence. To detach the cells from the bottom of the flask, cells were treated with trypsin.
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