Crl 2706

Two liver cell lines were used in this study: the non-tumorigenic cell line THLE-2 (ATCC® CRL2706™) and the hepatocellular carcinoma cell line HepG2 (ATCC® HB-8065™). Both cell lines were obtained from the American Type Culture Collection (ATCC).
Cells were maintained at 37°C in a humidified 5% CO₂ atmosphere. THLE2 cells were cultured in BEGM (CC-3170; Lonza™) plus the provided supplements and following the indications by ATCC, and supplemented, in addition, with 10 % FBS (S 0615; Invitrogen™, Life Technologies) and 1% antibiotic-antimycotic (15240062; Invitrogen™, Life Technologies). HepG2 cells were cultured in DMEM 1X (41965-039; Invitrogen™, Life Technologies) supplemented with 10% FBS and 1% antibiotic-antimycotic. Prior to any experiment, cells were synchronized under starvation (FBS free culture medium) overnight.

THLE-2 cells (derived from primary normal human liver cells) were purchased from American Type Culture Collection (CRL-2706™; Manassas, VA, USA). THLE-2 cells were cultured in LHC-8 medium (Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% fetal bovine serum (FBS; EURx, Gdańsk, Poland), 5 ng/mL epidermal growth factor (EGF; Thermo Fisher Scientific, Waltham, MA, USA), 70 ng/mL phosphoethanolamine (Sigma-Aldrich, Saint Louis, MO, USA), and 100 units/mL penicillin/streptomycin (Lonza, Basel, Switzerland). Cells were cultivated under standard conditions: 37 °C with 5% CO₂, and humidified atmosphere in the incubator on a T-75 cm² cell culture flask (Sigma-Aldrich, Saint Louis, MO, USA). To dissociate cell monolayer for the subculture, THLE-2 cells were washed with phosphate-buffered saline (PBS; Lonza, Basel, Switzerland) without calcium and magnesium, followed by treatment with trypsin 0.25%—EDTA in HBSS (Biosera, Nuaille, France)—for 4 min at 37 °C.