The number of viable cells in CCO1 and CCO2 was decreased only after treatment with both gemcitabine and LY294002 under Glu (−) condition, suggesting that the resistance of cholangiocarcinoma organoids to gemcitabine under glucose-free condition was dependent on AKT phosphorylation (Figure 4c). Here, AKT1 is linked to cholangiocarcinoma.