Culturing Mammalian Cell Lines for Experiments

An African green monkey kidney-derived cell line (VeroE6 cells) and a human hepatoma cell line (Huh7 cells) were maintained in Dulbecco’s modified Eagle’s medium (DMEM) (Fujifilm Wako), which was supplemented with penicillin and streptomycin sulfate (Thermo Fisher Scientific) and 5% fetal bovine serum (FBS) (Nichirei) for VeroE6 or 10% FBS for Huh7. Primary human hepatocytes (PHH) (PhoenixBio) were cultured with DMEM supplemented with 20 mM HEPES, 100 units/mL penicillin, 100 μg/mL streptomycin, 10% FBS, 44 mM NaHCO₃, 5 ng/mL epidermal growth factor (EGF), and 50 nM dexamethasone, as described previously (47 (link)). Primary human peripheral blood mononuclear cells (PBMC) (Lonza) were cultured with X-VIVO-15 (Lonza). The cells were then incubated under 5% CO₂ at 37°C.