HepG2 Cell Culture Protocol

HepG2 cells were obtained from the National Centre for Cell Science, Pune, India (local repository for the American Type Cell Culture (ATCC)), proliferated within two passage numbers, and stored in liquid nitrogen at passage number 17. Cells were cultured in high glucose (4.5 gm/L) DMEM cell culture media (Gibco, Evansville, IN, USA) supplemented with 10% FBS (HiMedia, Mumbai, India) and 100 U/mL penicillin/streptomycin (Thermo Fisher Scientific, Waltham, MA, USA) and maintained at 37 °C and 5% CO₂ [63 (link)]. Cells were passaged at 70% confluence and used for assays based on the number of cells required in each experiment. All experiments were conducted within 5 passages.