Huh28

CCA cell lines HuCCT1 and HuH28 were purchased from Japan Health Science Research Resources Bank, and CCA cell lines SNU-1196, SNU-1079, SNU-308, SNU-245, SNU-478 and SNU-869 were purchased from Korea Cell Line Bank. Human HEK293T was obtained from American Type Culture Collection (ATCC). All cell lines were cultured with RPMI-1640 medium (Invitrogen, USA) supplemented with 10% fetal bovine serum (Hyclone, USA) and 1% PenStrep (100 U/mL Penicilium and 100 μg/mL Streptomycin) in a humid atmosphere containing 5% CO₂ at 37 °C. Gemcitabine resistant cell lines HuCCT1-Gem and SNU-245-Gem were constructed by exposing to increasing dosages of Gemcitabine (Selleck Chemicals, USA) for 8 months, and then persistently cultured in medium containing 10 nmol/l Gemcitabine.

The human CCA cell line, HuCCT1 [34 (link)] and HuH28 [35 (link)] (Health Science Research Resources Bank, Osaka, Japan), which have significantly high and low expression of ITGB6 mRNA, respectively [7 (link)], was maintained in RPMI-1640 medium (Sigma–Aldrich, St. Louis, MO, USA) supplemented with 10% fetal bovine serum (FBS) (Sigma–Aldrich) and 1% antibiotic-antimycotic (Gibco, Dublin, Ireland) in a humidified atmosphere at 37 °C with 5% CO₂. These two cell lines were selected after previous experiments using five CCA cell lines with different characteristics reflecting heterogeneous cancer [7 (link)].