Hepatocellular Carcinoma Cell Lines

The HCC cell line Mahlavu was kindly provided by dr. Rengul Cetin-Atalay (Bilkent University, Ankara, Turkey) [65 –67 (link)] while Hep3B (ATCC no: HB-8064), HepG2 (ATCC no: HB-8065) SNU449 (ATCC no: CRL-2234) and SNU475 (ATCC no: CRL-2236) were obtained from ATCC. Mahlavu, Hep3B and HepG2 were maintained in DMEM medium supplemented with 10% FBS, 2 mM L-Glutamine, 0.1 mM NEAA, 100 U/ml penicillin and 100 μg/ml streptomycin. SNU449 and SNU475 cell lines were maintained in RPMI-1640 medium supplemented with 10% FBS, 2 mM L-Glutamine, 100 U/ml penicillin and 100 μg/ml streptomycin. For normoxic condition, all cells were cultured in a 37°C humidified incubator and an atmosphere of 5% CO₂ in air. For hypoxic condition, cells were cultured in a CO₂ incubator (Forma™ Series II Water Jacket CO₂, Thermo Scientific, USA) maintained at 94% N₂, 5% CO₂ and 1% O₂ for indicated times of treatments. Western blot analysis was performed by standard methods as described elsewhere [68 (link)–70 (link)].