Background: There have been lots of important discoveries in the research on embryonic stem (ES) cells; but few studies have been involving in the differentiation and differentiation ratio of hepatic stem cells. To provide a new source of hepatocytes in hepatocytes transplantation and liver tissue engineering, we investigated the differentiation and differentiation ratio of hepatocytes in the ES cells differentiating system in a certain length of time.
Methods: BALB/c ES cells from mice were cultured for differentiation in medium without leukemia inhibitory factor (LIF). During the period of differentiation of ES cells, dual immunofluorescence was performed to analyze the expression of hepatic proteins in cytoplasm such as alpha-fetoprotein (AFP) and albumin (ALB). The count of relative hepatocyte-like cells was analyzed by flow cytometry (FCM) and the hepatic differentiation ratio was obtained.
Results: The expression of AFP was first determined at day 7 and that of ALB at day 13. The expressions of AFP and ALB became stronger with the extention of the differentiation time. The hepatic differentiation ratio at day 13 and 21 was 5.5% and 10.4%, respectively.
Conclusion: ES cells could differentiate into hepatocytes, which should be a new source of hepatocytes in transplantation of hepatocytes and liver engineering.