Supporting culture medium

Human liver cancer cells; Huh7 and SNU449 were obtained from the Korean Cell Line Bank (Seoul, Korea) and maintained in Roswell Park MEMorial Institute medium (RPMI 1640; Welgene, Daegu, Korea). The human immortalized hepatocyte Fa2N-4 cell line were purchased from Xenotech (Lenexa, KS, USA), and maintained in serum-containing plating mediun (Xenotech) at first. After cell attachment in the plate, the medium was replaced with supporting culture medium (Xenotech). Human hepatic stellate cells (HSCs), LX2, was purchased from Merck Millipore (Darmstadt, Germany), and maintained Dulbecco`s modified Eagle`s medium (DMEM; Welgene) containing 2% fetal bovine serum (FBS; Gibco, Grand Island, NY, USA) and 1% penicillin–streptomycin (P/S; Gibco). One of HCC cell lines, HepG2 and WI38 human fibroblast cell line were obtained from ATCC (Manassas, VA, USA). These cell lines were maintained in minimum essential media (MEM; Welgene) supplemented with 10% FBS and 1% P/S. the Human umbilical vein endothelial cells (HUVECs) was purchased from PromoCells (Heidelberg, Germarny), and cultured in endothelial basal medium with supplementary reagents from PromoCells (Heidelberg, Germarny). All cells were maintained at 37 °C in a humidified incubator with 5% CO₂.

The Hep3B, Huh7, PLC/PRF/5, and SNU449 HCC cell lines were purchased from the Korean Cell Line Bank (Seoul, Korea). Huh7.5 cells were kindly provided by Dr. Marc Windisch (Institut Pasteur Korea, Gyeonggi-do, Korea), and Huh6 cells were obtained from Cell Bank Australia (Westmead, NSW, Australia). Human immortalized hepatocytes (Fa2N-4) were obtained from XenoTech (Lenexa, KS, USA). All cells were maintained at 37 °C in a humidified atmosphere of 5% CO₂. Dulbecco’s Modified Eagle’s Medium (Welgene, Daegu, Korea) was used to cultivate the Hep3B, Huh7.5, and Huh6 HCC cell lines, and Roswell Park Memorial Institute 1640 (RPMI1640) medium was used to cultivate the Huh7, PLC/PRF/5, and SNU449 HCC cell lines. DMEM and RPMI1640 media were supplemented with heat inactivated 10% fetal bovine serum (FBS; Gibco, Grand Island, NY, USA) and 1× penicillin-streptomycin (P/S; Gibco). Fa2N-4 cells were plated on collagen-coated plates (BD Biosciences, San Jose, CA, USA) in serum-containing plating medium (plating media; XenoTech), which was replaced with supporting culture medium (XenoTech) after cell attachment (approximately 3–6 h).