Culturing Human Liver Cell Lines

HCC cell lines, HCCLM3 and HepG2, were purchased from Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences (Shanghai, China). A normal liver epithelial cell line, LO2, and other two HCC cell lines, Hep3B and Huh7, were purchased from American Type Culture Collection (Manassas, VA, USA). All four HCC cell lines were cultured in DMEM (Invitrogen, Carlsbad, CA, USA) containing 10% fetal bovine serum (FBS; 10099141; Gibco, Carlsbad, CA, USA). The normal liver epithelial cell line LO2 was cultured in bronchial epithelial growth medium (ClonetIcs, Walkersville, MD, USA) containing epithelial growth factor (5 ng/mL) and phosphoethanolamine (70 ng/mL). Finally, the cells were cultured in a humidified incubator at 37°C with 5% CO₂ in air (thromo3111, Jinan Bisheng Medical Devices, Jinan, Shandong, China). The culture medium was replaced at the second day, and the cells were sub-cultured at the third to fourth day.³² (link)