Tfk 1

The human immortalized non-tumor cholangiocyte cell line (MMNK-1) and CCA cell lines (CCLP-1, HuCCT-1, KKU-055, KKU-100, KKU-213, KKU-214, RBE, and TFK-1) were obtained from the Japanese Collection of Research Bioresources (JCRB) Cell Bank (Osaka, Japan). The HuCCA-1 and RMCCA-1 CCA cell lines were developed from Thai patients with CCA (23 , 24 (link)). All cell lines were grown in Dulbecco’s modification of Eagle’s medium (DMEM; HyClone Laboratories, Logan, UT, USA) supplemented with 10% fetal bovine serum (Sigma-Aldrich, St Louis, MO, USA) and 1% Penicillin–Streptomycin (HyClone Laboratories) and were cultured in a humidified incubator at 37°C with 5% carbon dioxide. All cell lines were tested to be negative for mycoplasma contamination.

TFK1, RBE, SSP25, HuCCT1, HuCCA1, HuH28, KKU213A, KKU213B, KKU213C, KKU068, KKU131, KKU138, KKU055, KKU100, and YSCCC cell lines were obtained from the Japanese Collection of Research Bioresources (JCRB; Osaka, Japan). TFK1, RBE, SSP25, and YSCCC cells were maintained in Roswell Park Memorial Institute 1640 medium (RPMI 1640) (Gibco; Thermo Fisher Scientific, Waltham, MA, USA). HuCCA1 and KKU100 cells were maintained in Ham’s F-12 medium (Gibco; Thermo Fisher Scientific). HuCCT1, HuH28, KKU213A, KKU213B, KKU213C, KKU068, KKU131, KKU138, and KKU055 cells were maintained in Dulbecco’s Modified Eagle’s Medium (DMEM) (Gibco; Thermo Fisher Scientific). All cell lines were routinely tested for mycoplasma. For the construction of resistant cells, parental cells (KKU213C, TFK1, KKU068, and SSP25) were seeded in 12-well plates at 5 x 10⁴ cells/well in 1 milliliter (ml) of growth medium. Cells were treated with GEM/CIS using a stepwise dose-induction protocol (16 (link)).