Abstract: Objective To establish the method for isolation,culture,and differentiation induction of nestin-positive cells in fetal rat hepatic cells.Methods Hepatic cells were obtained from fetal rats by means of mechanical separation and hanging-drop culture,and after two days of primary culture,the medium was changed for further cell culture in the presence of 20% fetal bovine serum(containing glucose 25 ml/L,mycillin 100 U/ml,pH 7.6),10 mmol/L nicotinamide,1 mg/L insulin,affix N2,basic fibroblast growth factor,stem cell factor,epidermal growth factor and leukemia inhibitory factor.Result and conclusion Nestin-positive cells were obtained from fetal rat liver,which can differentiate into islet β cells after culture and expansion in vitro.