Culturing Hepatocellular Carcinoma and Primary Hepatocytes

Human Caucasian hepatocellular carcinoma cells HepG2 were purchased from European Collection of Cell Cultures (ECACC No. 85011430). Cells were cultured in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% of fetal bovine serum, 100 U/ml streptomycin, 100 µg/ml penicillin, 4 mM L-glutamine, 1% non-essential amino acids, and 1 mM sodium pyruvate. Cells were maintained at 37°C and 5% CO2 in a humidified incubator.
Primary human hepatocytes used in this study were obtained from two sources [19] (link): (i) from multiorgan donor HH52 (female; 60 years); the use of liver cells of donor HH52 was approved by “Ethical committee at the Faculty Hospital Olomouc”,and it was in accordance with Transplantation law #285/2002 Sb; “Ethical committee at the Faculty Hospital Olomouc” waived the authors from obtaining consent from the next of kin, regarding human hepatocytes obtained from liver donor HH52. (ii) long-term human hepatocytes in monolayer Batch HEP220770 (female; 35 years), Batch HEP220774 (female; 66 years) were purchased from Biopredic International (Biopredic International, Rennes, France). Cells were cultured in serum-free medium. Cultures were maintained at 37°C and 5% CO2 in a humidified incubator.