Mammalian cells used in this study were murine C2C12 cells (muscle, myoblast), hamster BHK-21 cells (kidney, fibroblast), human Huh7 cells (liver, hepatocellular carcinoma), human SVG-A cells (brain, astroglia) and human RD cells (muscle, rhabdomyosarcoma), chosen as we had previously shown that these cells efficiently supported CHIKV replication [14 (link)]. Mammalian cells were maintained in a humidified incubator at 37°C with 5% CO2 in complete Dulbeccos Modified Eagles Medium (DMEM) (Sigma) supplemented with 10% foetal calf serum (FBS) (Gibco) (or 20% FBS for C2C12 cells), 100 IU penicillin ml−1 and 100 μg ml−1 streptomycin (Gibco), 10% non-essential amino acids (NEAA) (Thermo Fisher Scientific) and 10 mM HEPES (Thermo Fisher Scientific). The mosquito cell line C6/36 (Ae. albopictus, embryonic) used in this study has a mutant Dcr2 gene that results in a defective RNAi response [15 (link)]. These cells were maintained at 28 °C in Leibovitz’s L-15 Medium (Thermo Fisher Scientific) supplemented with 10 % FBS, 100 IU penicillin ml−1 and 100 μg ml−1 streptomycin (Gibco), and 10% tryptose phosphate broth (Thermo Fisher Scientific).
Foetal calf serum fbs
Manufactured by Thermo Fisher Scientific
Sourced in United States
Foetal calf serum (FBS) is a cell culture supplement derived from the blood of bovine foetuses. It provides a complex mixture of proteins, growth factors, and other nutrients that support the growth and proliferation of cells in vitro.
Automatically generated - may contain errors
Lab products found in correlation
Leibovitz s l 15 medium, by Thermo Fisher Scientific (1 mentions)
Dulbecco s modified eagle s medium dmem, by Merck Group (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Non essential amino acids (neaa), by Thermo Fisher Scientific (1 mentions)
Hepes, by Thermo Fisher Scientific (1 mentions)
Tryptose phosphate broth, by Thermo Fisher Scientific (1 mentions)
Penicillin streptomycin, by Welgene (1 mentions)
3t3 l1 preadipocyte, by American Type Culture Collection (1 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions)
Dimethyl sulfoxide (dmso), by Thermo Fisher Scientific (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
3 protocols using foetal calf serum fbs
1
Cultivation of Diverse Mammalian Cell Lines
2
AZD1208 Modulates 3T3-L1 Adipocyte Differentiation
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3T3‐L1 preadipocytes (ATCC, Manassas, VA, USA) were grown up to the contact inhibition stage in DMEM supplemented with 10% heat‐inactivated foetal calf serum (FBS) (Gibco, Grand Island, NY, USA) and penicillin/streptomycin (Welgene, Daegu, Korea). Differentiation was induced by DMEM supplemented with 10% FBS plus a cocktail of hormones (MDI), containing 0.5 mmol L−1 IBMX (M), 0.5 μmol L−1 dexamethasone (D) and 5 μg/mL insulin (I) in the presence or absence of AZD1208. After 48 hours, the differentiation medium was replaced with DMEM supplemented with 10% FBS and 5 μg/mL insulin in the presence or absence of AZD1208 for 3 days. The cells were fed every other day with DMEM containing 10% FBS in the presence or absence of AZD1208 until day 8.
3
Cryopreservation of Human Blood Samples
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Peripheral blood (PB) and matched bone marrow‐derived aspirates (BM) were collected from patients with newly diagnosed AML (n = 15) and B‐ALL (BM, n = 11; PB = 12), which were provided by Queensland Children's Tumor Bank (Brisbane, Australia). Adult peripheral blood (APB) samples were collected from healthy individuals (n = 7). Umbilical cord blood (UCB; n = 8) was provided by the Queensland Cord Blood Bank at the Mater (Brisbane, Australia). All human sample donors gave written informed consent to sample acquisition following the Declaration of Helsinki, and the study has been approved by the Mater Human Research Ethics Committee (HREC/13/MHS/83, 22 310/1407AP and 1586 M) and ratified by the UQ HREC.
APB mononuclear cells (MNCs) were cryopreserved in 90% heat‐inactivated foetal calf serum (FBS; Gibco, Waltham, USA) and 10% dimethyl sulfoxide (DMSO; Sigma‐Aldrich, St. Louis, US). Cord blood MNCs were cryopreserved using 90% cord blood plasma and 10% DMSO solution. Cells were frozen at a maximum concentration of 50 × 106 cells mL−1 in 1‐mL cryovials. MNCs derived from AML and B‐ALL patient BM and PB samples were collected in EDTA vacutainer collection tubes and cryopreserved in 10% DMSO, 20% FBS and RPMI 1640 medium (Gibco). Cryovials were placed in NALGENE Mr.Frosty at −80°C for 12–24 h and subsequently relocated into long‐term −196°C liquid nitrogen storage.
APB mononuclear cells (MNCs) were cryopreserved in 90% heat‐inactivated foetal calf serum (FBS; Gibco, Waltham, USA) and 10% dimethyl sulfoxide (DMSO; Sigma‐Aldrich, St. Louis, US). Cord blood MNCs were cryopreserved using 90% cord blood plasma and 10% DMSO solution. Cells were frozen at a maximum concentration of 50 × 106 cells mL−1 in 1‐mL cryovials. MNCs derived from AML and B‐ALL patient BM and PB samples were collected in EDTA vacutainer collection tubes and cryopreserved in 10% DMSO, 20% FBS and RPMI 1640 medium (Gibco). Cryovials were placed in NALGENE Mr.Frosty at −80°C for 12–24 h and subsequently relocated into long‐term −196°C liquid nitrogen storage.
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