Obeticholic acid (OCA) was provided by Intercept Pharmaceuticals, Inc. San Diego, USA, and was prepared as stock solution in dimethyl sulfoxide (DMSO, CAS Number 67-68-5, Sigma-Aldrich, Milan, Italy) and then diluted (1:105) in culture medium at the desired final concentration; the same amount of DMSO was added to controls. Stock solutions of OCA were freshly prepared every 15 days.
Gemcitabine hydrochloride (Sigma-Aldrich, CAS Number: 95058-81-4) was prepared as a stock solution in water and added in the culture media of iCCA cells after dilution (1:104). Cisplatin (Sigma-Aldrich, CAS Number: 15663-27-1) was prepared as a stock solution in DMSO and added to cell cultures, after appropriate dilution (1: 105) in H69 medium. We used the concentration of Gemcitabine (10 μM) and Cisplatin (20 μM) previously demonstrated to exert maximal inhibitory effects on cell proliferation in the same primary human iCCA cell cultures used in the present study [19 (link)].
Chenodeoxycholic acid (CDCA) and ursodeoxycholic acid (UDCA) were purchased from Sigma-Aldrich, diluted in DMSO and added into culture medium after dilution to reach the desired final concentration. The same amount of DMSO was added to controls.
During the experiments, the cell medium was refreshed every 3 days.
Gemcitabine hydrochloride (Sigma-Aldrich, CAS Number: 95058-81-4) was prepared as a stock solution in water and added in the culture media of iCCA cells after dilution (1:104). Cisplatin (Sigma-Aldrich, CAS Number: 15663-27-1) was prepared as a stock solution in DMSO and added to cell cultures, after appropriate dilution (1: 105) in H69 medium. We used the concentration of Gemcitabine (10 μM) and Cisplatin (20 μM) previously demonstrated to exert maximal inhibitory effects on cell proliferation in the same primary human iCCA cell cultures used in the present study [19 (link)].
Chenodeoxycholic acid (CDCA) and ursodeoxycholic acid (UDCA) were purchased from Sigma-Aldrich, diluted in DMSO and added into culture medium after dilution to reach the desired final concentration. The same amount of DMSO was added to controls.
During the experiments, the cell medium was refreshed every 3 days.
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