| Objective:To establish a stable and reliable hepatic oval cells isolation method and build a solid foundation for the subsequent experiment research.Methods:12 cases of the transgenic C57BL/6 mice, which were carried green fluorescent protein(GFP),10 of them were incorporated 3,5-diethoxycarbonyl-1,4-dihydro-collidine (DDC) in a standard food at a concentration of 0.1% to activate the oval cells proliferation,then the nonparenchymal cells in the liver were separated by in situ perfusion,finally we used Sca-1 antibody in conjunction with magnetic activated cell sorting (MACS) to separate the Sca-1+ cells.Result:The pathological section shows conspicuous proliferative reactions in liver tissue after 6 weeks of DDC diet. We get 106/ml non nonparenchymal cells from each mouse.The cells have 10-15μm in diameter, cobblestones overlook, big nucleus, low endochylema under the electronic microscope. The results of identification prove that the Sca-1+ cells we separated had a morphology in accordance with hepatic oval cells, and expressed the surface markers of OV-6 and AFP.Conclusion:The DDC diet protocol for mouse oval cells is reliable,and the MACS can be a efficient method to isolate the hepatic oval cells. Objective:To investigate the feasibility of hepatic oval cells to treat cirrhosis in mice.Methods:40 casese of SPF C57BL/6A mice were injected subcutaneously with CC14 to establish experimental liver cirrhosis model.Then the cirrhosis mice model were divided into two groups.The mice of one group were transplanted with the hepatic oval cells by spleen injection(HOC group),and the other one were injected with saline instead as a control group (NS group). At the meanwhile, the mice in each group were divided into two sub group, they were group of keeping injection of CCl4 (A group)and group of stopping CCl4 injection after transplantation(B group). Four weeks later, we investigate the liver function index, the hydroxyproline and collagen contents and the histopathological change among the groups to evaluate the effect of hepatic oval cells transplantation.Result:It shows that the majority of hepatic oval cells, which were transplanted transpleen into the mice, were planted in the portal area under. The levels of ALT and AST were conspicuous higher in NSA group compared with that in HOCA (p<0.05). TP and ALB increased after transplantation but didn't show significance in statistics.The examination in pathology showed that cirrhosis relieved in HOCA group and got severe in NSA group.The results demonstrate that the function index and histopathological change were improved in the experimental group, especially in the group of ceasing CC14 injection after transplantation.Conclusion:After transplanted into the liver by spleen injection, the hepatic oval cells could plant in the portal area, improve the liver fuction, and alleviate the cirrhosis level. Objective:To compare the effectiveness of hepatic oval cells and bone marrow mesenchymal stem cells to treat cirrhosis in mice.Methods:The mice were injected subcutaneously with CC14 to establish experimental liver cirrhosis.Then 60 cases of the cirrhosis mice were divided into three groups.The experimental was transplanted the hepatic oval cells by spleen injection(HOC), another group was transplanted with bone marrow mesenchymal stem cells(MSC), and the last group was injected saline instead as a blank control group(NS). In the while, the mice in each group were divided into two subset group, they were group of keeping injection of CCl4 (A group) and the group of stopping CCL4 injection (B group). Four weeks later, we investigate the liver function index, the hydroxyproline and collagen contents and the histopathological change among the groups to evaluate the effect of hepatic oval cells and bone marrow mesenchymal stem cells transplantation.Result:85 cases the transgenic C57BL/6 mice,6 of them were sacrificed for bone marrow mesenchymal stem cells,79 of them were used for making cirrhosis mice model and succeeded in 64 cases (81%). After transplantation with hepatic oval cells and bone marrow mesenchymal stem cells, The hydroxyproline and collagen fibers in liver tissue were decrease in HOC group (p<0.05), The examination in pathology showed that liver cirrhosis level had improved in every A groups with different degree (HOCB |
PDF Full Text Request