Objective: To investigate the method of isolation and culture of hepatic stellate cells for studying the cellular mechanisms of hepatic fibrosis. Methods: The isolation of normal rat hepatic stellate cells was established with collagenase in situ liver recirculating perfusion.Hepatic stellate cells were purified by using density gradient centrifugation with 18% Nycodenz. Results:The amount of hepatic stellate cells obtained were 1×107～3×107 cells per liver with a purity of more than 90% and the viability of more than 96%. Conclusion: The result showed that the methods of the isolation and culture of rat hepatic setllate cells were economical and steady.