Study On Directed Differentiation Of Human Umbilical Cord Blood Mesenchymal Stem Cells Into Hepatocyte-Like Cells In Vitro And Treatment Of Liver Injury With Them

Objective:To explore the effection of transplanting human umbilical cord blood MSCs into rats injuried by carbon tetrachloride in the liver.Methods:Normal parturients' cord blood were collected.Mscs from human umbilical cord blood were isolated by density gradient centrifugation and adherence methods,and then to be purified and frozen.Expression of MSCs' surface antigen CD34 and CD44 were detected by flow cytometry.The P3 Mscs were induced by HGF and FGH-4 into hepatocyte-like cells after purification and cultivation.The markers of hepatic linear cells were examined by immunocytochemistry and periodic acid-schiff (PAS).After that,using transmission electron microscope to observe the ultrastructure of cells with HGF and FGH-4 induction.Finaly,transplanting the MSCs and the hepatocyte-like cells into rats with chronic liver injury,then observing the liver pathology.Results:1.the density gradient centrifugation and adherence methods are fit for the separation and growth of umbilical cord blood MSCs.2.the induced cells after purificating and going down to the third generation shows ultrastructure of hepatocyte-like cells.3.Whereas in the groups without HGF and FGH-4 induction,the cells did not express hepatocellular markers:ALB,hepatin but AFP in 0 day.Whereas in the groups with HGF and FGH-4 induction,the cells express hepatocellular markers: ALB,hepatin and AFP.The third generation cells express surface antigen CD44 but not CD34.Both the MSCs and the hepatocyte-like cells can alleviate the liver injury of rats,and the hepatocyte-like cells transcend the MSCs.Conclusion:1.human umbilical cord blood MSCs can be isolated successfully with density gradient centrifugation and can be cultured successfully with adherence methods.2.human umbilical cord blood MSCs could be induced into the hepatocyte-like cells by HGF and FGH-4 in vitro.3.40%carbon tetrachloride can make the model of rat chronic liver injury successfully.4.Both the MSCs and the hepatocyte-like cells can alleviate the liver injury of rats,and the hepatocyte-like cells transcend the MSCs.5. MSCs can be preserved by frozen for long.