Objective To explore the techniques of isolation and in vitro culture of hepatic progenitor cells of normal rat liver. Methods According to the cell surface marker CD90.1, hepatic progenitor cells of normal Wistar rats were isolated by immunomagnetic cell sorting, cultured in vitro, and induced to differentiate. Results The percentage of CD90.1 positive cells in non-parenchymal cells was (0.32±0.03) %. The flow cytometric cell sorting showed that (98.26±1.37) % of isolated cells were CD90.1 positive cells. Fresh isolated CD90.1 positive cells were strong positive for CK-19 and negative for HNF-4a. After 8 weeks of culture, these cells were negative for CK-19 and positive for HNF-4a. Conclusion Hepatic progenitor cells exist in normal rat livers and can be differentiated toward hepatocytes.