Abstract: Objective To establish the implantation model of human in situ hilar cholangiocarcinoma cell line in nude mice.Methods Cultured human hilar cholangiocarcinoma cell line QBC939 was injected into the space between the hilar bile duct and portal vein of nude mice with a micro-injector.Anatomical and pathological features of the mice were detected 15d after the implantation model of human in situ hilar cholangiocarcinoma cell line was established.Results Suspension of 100ml of hilar cholangiocarcinoma cells at the concentration of 1×106/ml was injected into the space between the hilar bile duct and portal vein of each nude mouse.The successful rate was 100% 15d after the implantation model of human in situ hilar cholangiocarcinoma cell line.Histological examination showed typical tumor tissues in mice.Conclusion An implantation model of human in situ hilar cholangiocarcinoma can be established in nude mice.