K4000

The HCC cell lines ;SNU449, SNU475, SNU398, SNU898, Huh7, HepG2, Hep3B and PLC/PRF/5 were purchased from the Korean Cell Line Bank. Huh6 cells were kindly provided by Dr. Ralf Bartenschlager (University of Heidelberg, Germany). All HCC cells were maintained in RPMI (Welgene, Korea) or Dulbecco’s Modified Eagle Medium (DMEM; Welgene, Korea) containing 10% fetal bovine serum (FBS; Gibco, Grand Island, NY, USA) and 1% penicillin/streptomycin solution (p/s; Gibco, Grand Island, NY, USA). Fa2N-4, a human immortalized hepatocyte cell line, was obtained from Xenotech (Lenexa, KS, USA) and first cultured in serum-containing plating medium (K4000; Xenotech). Miha. Cells were kindly provided by Dr. Kim. K. M. from ASAN medical center. To produce tumor spheroids, HCC cells seeded at a density of 6 × 10³ cells/well in 96-well round-bottom ULA microplates (Corning Life Sciences, Corning, NY, USA). All cells were maintained in a 5% CO₂ humidified incubator at 37 °C.

Huh7, Hep3B, PLC/PRF/5 and HepG2 cells (human HCC lines) were obtained from the Korean Cell Line Bank. Human HCC cell line Huh6 was kindly provided by Dr. Ralf Bartenschlager (University of Heidelberg, Germany) and Fa2N-4 cells (human immortalized hepatocyte cell line) were purchased from Xenotech (Lenexa, KS, USA). HCC cell lines were cultured in Dulbecco’s minimal essential medium (DMEM; Welgene, Korea, LM001-05) supplemented with heat-inactivated 10% fetal bovine serum (FBS; Gibco, Gaitherburg, MD, USA) and 100U/ml Penicillin and 100 μg/ml Streptomycin (Gibco) at humidified 37 °C incubator under 5% CO₂. Fa2N-4 cells were plated in collagen-coated plates. After cell attachment (approximately 3 ~ 6 h), serum-containing plating medium (XenoTech, K4000) was replaced with supporting culture medium (XenoTech, K4100.X).