ABSTRACT
Objective:To observe the conditions of isolation,purification and culture of mesenchymal stem cells(MSCs) in vitro derived from human umbilical cord,and to observe the repairing effect on experimental hepatic fibrosis in rats by transplantation with MSCs only or MSCs with IL-10.Methods:The MSCs derived from human umbilical cord were isolated,purified and cultivated in vitro.The model of hepatic fibrosis induced by CCl4 was established.After the model of hepatic fibrosis was succeeded,the rats were divided into three groups:model group,MSCs transplantation group,MSCs transplantation with IL-10 group.MSCs transplantation group was given 1×106 MSCs by the tail vein injection at the first 1 weeks,2 weeks and 3 weeks.MSCs transplantation with IL-10 group not only was given MSCs the same with MSCs transplantation group, but also was injected of IL-10 ( 4 μg/kg ) in abdomen, 4 times a week.After 4 weeks,the rats were executed,rats blood was collected to observe the changes of liver function and the index of liver fibro-sis.The liver HE staining was carried out to observe the changes of liver pathology.Expression of MMP-2 mRNA was detected by RT-PCR method,expression of TGF-βprotein was detected by Western blot method.Results:The rat liver function and the index of liver fibrosis improved obviously with a significant difference between model group , MSCs transplantation group and MSCs transplantation with IL-10 group after transplanting,MSCs with IL-10 was better than MSCs only.Liver tissue of rats with hematoxylin eosin staining suggested,liver fibrosis was obviously improved,and the MSCs transplantation with IL-10 group was best.MMP-2 mRNA expression was significantly higher between normal group and model group,MSCs transplantation group with a significant difference (P0.05).TGF-βprotein expression of other groups was higher than normal group with a significant (P<0.05),and MSCs transplantation with IL-10 group was the largest decrease than normal group.Conclusion:MSCs only or MSCs with IL-10 transplantation can improve the biochemical characteristics of rat peripheral blood and liver histological structure,and with IL-10 wan better than MSCs only.MSCs and IL-10 play a role in the treatment of liver fibrosis through lowering the expression of MMP-2 and the expression of TGF-βprotein.
ABSTRACT
BACKGROUND:Recent studies verified that other tissues-derived stem cells can be homing to the liver, possibly participate in the regeneration of liver tissues, which provides new hope for stem cells in treatment of liver disease. OBJECTIVE:To observe isolation and culture of human umbilical cord-derived mesenchymal stem cells, to observe the repairing effect of umbilical cord-derived mesenchymal stem celltransplantation on hepatic fibrosis, and then to provide a reliable theoretical basis for further clinical application of umbilical cord-derived mesenchymal stem cells. METHODS:Human umbilical cord-derived mesenchymal stem cells were isolated and purified by natural adherent method, and then cultured and amplified in vitro. A rat model of hepatic fibrosis was prepared using subcutaneous multi-point injection of CCl4. A total of 22 rat models were randomly assigned to model injury group (n=11) and celltransplantation group (n=11). At 1, 2 and 3 weeks after model induction, the rats in the celltransplantation group were treated with 1×106 umbilical cord-derived mesenchymal stem cells via caudal vein. Four weeks later, the rats were sacrificed. Blood of rats was col ected from each group to detect liver functions. The liver was removed to receive hematoxylin-eosin staining, and pathological changes were observed. The number and distribution of Kupffer’s cells were observed using immunohistochemistry. The localization of umbilical cord-derived mesenchymal stem cells from treatment group was observed using immunohistochemistry.RESULTS AND CONCLUSION:After umbilical cord-derived mesenchymal stem cells were infused into rats with cirrhosis via caudal vein, liver function was significantly improved, which showed significant differences as compared with the control group (P<0.05). Hematoxylin-eosin staining revealed that hepatic fibrosis was apparently improved. Immunohistochemistry results demonstrated that the number of Kupffer’s cells was obviously reduced, and BrdU-labeled umbilical cord-derived mesenchymal stem cells were visible in rat liver of the treatment group using anti-BrdU antibody. These results suggested that umbilical cord-derived mesenchymal stem celltransplantation could improve biochemical characteristics of rat peripheral blood and histological structure of liver.