Cell culture

Thai patient-derived CCA cell lines, KKU-055 (JCRB1551) [³⁴], KKU-100 (JCRB1568) [³⁶], KKU-213 (previously known as KKU-M213) (JCRB1557) [⁴¹] and HuCCA-1 (JCRB1657) [³⁵], as well as immortalized cholangiocyte cell line MMNK-1 (JCRB1554) [⁴²] from the Japanese Collection of Research Cell Bank (JCRB) were maintained in a humidified 5% CO₂ incubator at 37°C in HAM’s F-12 media (GIBCO, Grand Island, NY, USA) containing 15 mM HEPES, 14 mM NaHCO₃, 100 U/ml penicillin G (GIBCO), 100 U/ml streptomycin (GIBCO), and 10% heat-inactivated fetal bovine serum (FBS) (GIBCO). KKU-055 and KKU-100 were developed from patients with poorly differentiated adenocarcinoma and the latter appears to be multidrug resistant cell lines [³⁴]. Basal proliferation, migration and invasion abilities of HuCCA-1 were relatively low when compared to that of KKU-213 [^21,35,43].