Hucct1 cell line

The HuCCT-1 cell line, derived from human Cholangiocarcinoma (CCA), was obtained from the Japanese Collection of Research Bioresources (JCRB) Cell Bank in Osaka, Japan. The RMCCA-1 cell line, also a human CCA cell line, was developed from Thai CCA patients [43 (link)]. Both HuCCT-1 and RMCCA-1 cells were cultured in HAM’s F-12 medium (HyClone Laboratories, Logan, UT, USA) supplemented with 10% fetal bovine serum (Sigma, St Louis, MO, USA) and 1% Penicillin-Streptomycin (HyClone Laboratories, Logan, UT, USA). All cell lines were maintained in a humidified incubator at 37 °C with 5% CO₂. Furthermore, rigorous testing confirmed that there was no mycoplasma contamination in any of the cell lines.

Patient sample acquisition was performed under an Institutional Review Board (IRB) protocol approved at California Pacific Medical Center in accordance with relevant guidelines and regulations. Informed consent was obtained from the patient (Male, 58yrs old, tumor moderately differentiated, and stage IV) in accordance with approved institutional guidelines. PDX generation, STR analysis and PDXC culture conditions are previously described by our group^{30 (link)}. The human CCA HuCCT1 cell line was purchased from the Japanese Collection of Research Bioresources Cell Bank (JCRB, Japan) and KMCH was kindly provided by Dr. Gregory Gores (Mayo Clinic, MN). HuCCT1 and KMCH were grown in RPMI (Thermofisher Scientific, South San Francisco, CA) with 5% fetal bovine serum (JR Scientific, Woodland, CA) and 1 × penicillin/streptomycin (Thermofisher Scientific) at 37 °C in a 5% CO₂ incubator. Cell lines tested negative for mycoplasma contamination using MycoFluor Mycoplasma Detection Kit (Thermofisher Scientific) following manufacturer’s instructions. Dinaciclib, gemcitabine and palbociclib were purchased from Selleck chemicals (Houston, TX).