The invention relates to an adult human hepatogenic stem cell line HN, a preparation method thereof and an application. The human hepatogenic stem cell line is collected at the China General Microbiological Culture Collection Center, the collection address is No. 3, first yard, Beichen West Road, Chaoyang District, Beijing, the collection date is May 15, 2017, and the collection number is CGMCC No. 13844. The human hepatogenic stem cell line is used for in vitro and vivo directional induction of liver cells and hepatic duct epithelial cells, preparation of biological artificial livers, resistance of acute hepatic failure, chronic hepatic failure and alcoholic fatty livers, inhibition of liver fibrosis and anti-tumor effects. The cells have the biological characteristics that the cells have typical epithelial cell morphology and strong multiplication capacity, grow to achieve contact inhibition and density inhibition, bidirectionally express liver cell specific marker albumin, cytokeratin 8 and biliary epithelial cell specific marker cytokeratin 7 and 9, have various functions of the liver cells, have multi-directional differentiation potential and avoid tumorigenicity.