Wi 38 cells

WI-38 cells, which are human normal embryonic lung-derived diploid fibroblasts (population doubling level, 23), were purchased from the Korean Cell Line Bank and grown in DMEM containing 10% FBS and 1% penicillin/streptomycin until reaching confluence.

Twelve biotin receptor-positive cell lines; human lung carcinoma cells (A549), human cervical cancer cells (HeLa), human breast cancer cells (MCF7, MDA-M231), human liver cancer cells (HepG2, Huh7, Hep3B), human prostate cancer cells (Du145, PC3), human gastric cancer cells (NCI-N87, AGS), human pancreatic cancer cell (Panc-1), and a biotin receptor-negative cell: human normal embryonal kidney epithelial cell (293T) were purchased from the Korean Cell Line Bank (Seoul, Republic of Korea). Two more biotin receptor-negative normal cell lines, human normal fibroblast cells obtained from fetal lung (WI-38 cells) or neonatal foreskin (BJ) cells were purchased from the Korean Cell Line Bank (Seoul, Republic of Korea) or Modern Cell & Tissue Technologies (MCTT, Seoul, Republic of Korea). The cells were cultured in either Roswell Park Memorial Institute medium (RPMI-1640, GIBCO BRL, Grand Island, NY, USA) or Dulbecco’s Modified Eagle’s Medium (DMEM, GIBCO BRL) supplemented with 10% fetal bovine serum (FBS, GIBCO), and 1% penicillin and streptomycin (GIBCO), at 37 °C in a humidified atmosphere containing 5% of CO₂. When the cell density reached 70–80% of confluence, subculturing was considered complete. The medium was changed approximately every 3 to 4 days.

LX2 cells (human hepatic stellate cells; HSCs) were provided by Dr. Haeng Ran Seo (Institute Pasteur Korea). HUVEC cells (human umbilical vein endothelial cells) were obtained from Lonza (Basel, Switzerland). WI38 cells (human fibroblast cell line) were obtained from the Korean Cell Line Bank. The cells were maintained at 37 °C in a humidified atmosphere (5% CO₂/95% air). LX2 cells were cultured in Dulbecco’s Modified Eagle Medium (DMEM; Gibco, Grand Island, NY, USA) supplemented with heat-inactivated 10% fetal bovine serum (FBS; Gibco, Grand Island, NY, USA) and 1× penicillin (Welgene, Gyeongsan, Korea) (Complete media). HUVEC cells were maintained in Medium 200 (Gibco, USA) supplemented with 1× penicillin (Welgene, Gyeongsan, Korea), 1× LSGS (Gibco, USA) and heat-inactivated 10% FBS. For WI38 cells, Roswell Park Memorial Institute medium (RPMI 1640; Gibco) supplemented with 1× penicillin (Welgene, Gyeongsan, Korea), and 10% heat-inactivated FBS was used. Primary HCC cells were maintained in DMEM/F12 (Gibco, USA) supplemented with 1× penicillin (Welgene, Gyeongsan, Korea), 1× GlutaMmax (Gibco, USA), 1× Primocin (Invitrogen, Carlsbad, CA, USA), and 10% heat-inactivated FBS.