Hepg2

The human HCC cell lines Huh7 and HepG2 were purchased from the Cell Bank of the Type Culture Collection of the Chinese Academy of Sciences (Shanghai, China). Cells were cultured in RPMI-1640 medium (Gibco, Carlsbad, CA, USA) supplemented with 10% fetal bovine serum (Gibco). The cells were cultured at 37°C in a humidified incubator with 5% CO₂. To obtain the CD133⁺ and CD133^- Huh7 and HepG2 cells, the cultured cells were stained with anti-CD133-FITC antibody (Miltenyi Biotec, Germany) for 20 min at room temperature. The population of CD133⁺ and CD133^- Huh7 and HepG2 cells were analyzed and sorted by using the fluorescent-activated cell sorting equipment (Beckman Coulter, USA).

The HCC cell lines HepG2 and Huh7 were purchased from the Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences (Shanghai, China) and maintained in RPMI-1640 (Gibco, USA) supplemented with 10% fetal bovine serum (FBS, Gibco) at 37°C in a humidified 5% CO₂ incubator. For separation between CD133⁺ HCC cells and CD133^- HCC cells, HepG2 and Huh7 cells were incubated with fluorescein isothiocyanate (FITC) conjugated CD133 antibody (Miltenyi Biotec, Germany) for 20 min at room temperature. Subsequently, CD133⁺ and CD133^- HepG2 and Huh7 cells were sorted by FACS vantage (FACSCALIBUR, BD Biosciences, USA).