Objective To investigate biological characteristic of hepatic progenitor cells from fetal liver of rat on ED13.5 days and establish the foundation for liver cell transplantation. Methods Fetal liver cells were isolated by enzymatic digestion. Different influence on cellular biologic action was observed under various cultured conditions and liver specific markers of cells were detected by immunocytochemistry. Results Fetal liver epithelial progenitor cells proliferated on primary rat embryonic fibroblast (PREF) feeder better than non-feeder and were kept undifferentiation. Cell differentiation marks suggested that fetal liver epithelial progenitor cells contain bipotential cells on ED13.5 d.Conclusion There retains hepatic stem cells in rat fetal liver on ED 13.5 d and PREF can be used for amplification of fetal liver epithelial progenitor cells in vitro.