HepG2 Cell Culture and MTT Assay

Human hepatoma HepG2 cell line was purchased from the National cell bank of Iran (NCBI, Pasteur Institute, Tehran). Cells were cultured in RPMI 1640 medium supplemented with 10% FBS, 2mM Gln, penicillin (100 U/mL), and streptomycin (100 μg/mL) in humidified 5% CO2, the atmosphere at 37°C temperature. For ChiNH, Q, and ChiNH/Q treatment, cells were seeded into six-well plates at a density of 3 × 10⁴ cells and allowed to attach for 24 h. The viability of HepG2 cells was determined by MTT test using 5 different concentrations of 10, 50, 100, 500 and 1000 µg/mL of ChiNH, Q and ChiNH/Q for 48h based on the method described by Rashidipour et al.⁵² All experiments were performed in triplicate and the data were expressed as mean ± SD.