Ham f 12 nutrient mixture ham f 12

CCA cells (HuCCA-1 and RMCCA-1) established from CCA tissue fragments of Thai patients were tested for anti-migration effects of SG. HuCCA-1 is derived from a patient with intrahepatic bile duct CCA (Sirisinha et al., 1991) . RMCCA-1 is derived from a patient with peripheral CCA (Rattanasinganchan et al., 2006) . CCA cells were grown in 6-well plates overnight before treatments. They were cultured at 37 °C, 5% CO 2 in Ham F-12 nutrient mixture (Ham F-12) (Gibco Invitrogen, USA) supplemented with 1 mM L-glutamine, 1.17 g/l sodium bicarbonate (NaHCO 3 ), 10% of FBS (Sigma Aldrich, USA) and penicillin (100 units/ml) plus streptomycin (100 µg/ml) (Wiscent Inc. P.O., Canada).

CCA cells (HuCCA-1, RMCCA-1 and KKU-M213) established from CCA tissue fragments of Thai patients were tested for an anti-proliferation effect of SG. HuCCA-1 (Sirisinha et al., 1991) and KKU-M213 are derived from a patient with intrahepatic bile duct CCA. RMCCA-1 is derived from a patient with peripheral CCA (Rattanasinganchan et al., 2006) . They were cultured at 37°C, 5% CO 2 in Ham F-12 nutrient mixture (Ham F-12) (Gibco Invitrogen, USA) containing 1.17 g/L sodium bicarbonate (NaHCO 3 ), 5% FBS (Sigma Aldrich, USA) and penicillin (100 units/mL) plus streptomycin (100 µg/mL) (Wiscent Inc. P.O., Canada).