Experimental Study On Hepatic Oval Cells To Establish Proliferation Model And Isolate In Vitro In Adult Rat

Background: Hepatic dysfunction is a common syndrome of all kinds of end-stage liver disease, and nowadays liver transplantation is the unique therapy. But for the lack of donor liver, the application of liver transplantation is limited. The application of hepatocyte transplantation is becoming a bridge when a patient is waiting for liver transplantation. However, the source of hypatocyte is much limited. The appearance of hepatic oval cells(HOCs) just solves the problem. HOCs is a kind of stem cells which can differentiate into hepatocytes and cholangioepithelial cells.Objective: To seek for the best way of establishing a model of hepatic oval cells(HOC) proliferation and study the better methods of isolating HOC in vitro.Methods: 30 male Wistar rats weight(150±10 )g were randomly divided into control group and model group . HOC proliferation model was established with 2-AAF+2/3PH(2-acetylaminofluorene+two thirds partial hepatectomy). The model group was administrated 15mg/kg/d of 2-AAF by garage, the control group was administrated the same dose of normal sodium ,then a standard PH were performed on day 5, the same dose was still administrated as preoperation, the biopsies were adopted and identified by immunohistochemical staining at 10～14d postoperation for observation of the proliferation of HOC. The remarkably proliferated HOC were isolated by collagenase perfusion method and percoll gradient centrifugation, which were identified by optic microscope.Results: Pathologic examination of the rat liver revealed that the HOC were induced in rats with administered 2-AAF combined with 2/3PH. but HOC of rat liver in the control group were not obviously observated. HOC were positived with CK19 andC-kit, which were effectively isolated by Collagenase perfusion method and percollgradient centrifugation.Conclusions: The proliferation model of HOC was ideally established with2-AAF+2/3PH, Collagenase perfusion method and percoll gradient centrifugation canbe used to isolate HOC effectively, which is a effective methods to isolate adult liverstem cells.