Abstract: Objective To study the expression of nestin in OCT4-mediated dedifferentiation of hepatocytes and its significance. Methods Mouse primary hepatocytes were isolated and cultured in vitro. Hepatocytes were infected with packaged and concentrated OCT4- lentivirus. Morphology of hepatocytes was observed. Expressions of nestin, ALB, AFP, and FOXA2 were detected by RTPCR and quantitatively analyzed. Results After the hepatocytes were infected with OCT4-lentivirus, the intracellular particles were gradually released from hepatocytes and the hepatocytes began to proliferate from day 7, the nestin increased its expression from day 3 to day 7 and began to decrease from day 12 when the ALB reduced it expression while the AFP and FOXA2 increased their expressions. The expressions of these genes were hardly detected on day18 after the hepatocytes were infected with OCT4-lentivirus. Conclusion Nestin is expressed with the dedifferentiation and proliferation of hepatocytes.