Isolation,Purity and Identification of Hepatocytes in Rat Normal Liver and Regenerating Liver

Rat 2/3 hepatectomy model was made by the method of Higgins et al.,hepatic cells were scattered by two-step perfusion,and hepatocytes were isolated and pruified by density gradient centrifugation with 60 % percoll. Immunocytochemistry method was used to qualitify and localize albumin(ALB) and glucose-6-phosphatase(G-6-P) in liver tissue,the dispersed hepatic cells,and the purified hepatoctyes. The exrpressions of alb and G-6-P were quantified using RT-PCR. The results primarily showed that ALB/G-6-P postive hepatocytes account up more than 96 % of the total hepatocytes;mRNA levels of ALB and G-6-P in the purified hepatocytes were stable in the Purificl hepatocytes of rat regenerating liver,and also was the content of the corresponding proteins,indicating the modified method for hepatocytes purification in this study has the adventage of high hepatocyte harvest,high purity and survival rate.