The human liver cancer cell line, HepG2, was obtained from the European Collection of Authenticated Cell Cultures (cat. no. 85011430). The cells were seeded into 75-cm2 T-flasks (Falcon; Corning Life Sciences) in DMEM (Gibco; Thermo Fisher Scientific, Inc.) supplemented with 10% (vol/vol) fetal bovine serum (FBS; Gibco; Thermo Fisher Scientific, Inc.), 4 mM L-glutamine (Gibco; Thermo Fisher Scientific, Inc.) and 100 U/ml penicillin-streptomycin with fungizone (Gibco; Thermo Fisher Scientific, Inc.), and incubated in a 5% CO2 atmosphere at 37°C. The cultures were trypsinized and sub-cultured once a week.
75 cm2 t flasks
Manufactured by BD
The 75-cm2 T-flasks are cell culture vessels designed for the in vitro cultivation of cells. They provide a surface area of 75 square centimeters for cell growth and proliferation.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Fungizone, by Thermo Fisher Scientific (1 mentions)
Hepg2, by European Collection of Authenticated Cell Cultures (1 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)
L glutamine, by Thermo Fisher Scientific (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Cryostor cs10 freezing media, by BioLife Solutions (1 mentions)
2 protocols using 75 cm2 t flasks
1
HepG2 Cell Line Cultivation Protocol
2
Expansion and Cryopreservation of eCB-MSCs
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For the static culture, the eCB-MSCs were expanded in 75cm2 T-flasks (Falcon Cat#: 353136) at an inoculation density of 5000 cells/cm2, with 12 mL of media, in a humidified incubator (37 °C and 5% CO2 in ambient air). Once the cells neared confluence (~ 80%), they were harvested by exposing the cells to 0.25% Trypsin for 5 min in a humidified incubator (37 °C and 5% CO2 in ambient air), followed by deactivation of the Trypsin using FBS containing media. The cells were then enumerated on a haemocytometer using 0.1% Trypan Blue exclusion, and either passaged onto new T-flasks, inoculated on microcarrier beads within bioreactors, or cryopreserved in Cryostor CS10 freezing media (BioLife Solutions Cat # 210102) for future cell characterization.
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