Abstract:

BACKGROUND: Hepatocyte transplantation as a role of bridge can help the liver failure patients to pass through the liver failure phase and improve patient survival rate and prognosis. 
OBJECTIVE: To research the improvement of method and the influencing factors of primary hepatocytes isolation technology in Sprague Dawley rats by Seglen modified in situ perfusion method, and to evaluate the treatment effects of primary hepatocytes transplantation on rats with acute hepatic failure.
METHODS: Rats’ hepatocytes were isolated by the two step collagenese digestion, and acute hepatic failure was induced by D-galactosamine in rats. After 24 hours, the acute hepatic failure rats were divided into two groups randomly. The rats in the transplantation group received 2×107 hepatocytes through intrasplenic injection, the rats in the control group received 0.4 mL Hank’s solution by the same way. The survival rate, level of transaminase and total bilirubin in serum, secretion of albumin and the distribution of hepatocytes in spleen were observed at different time points.
RESULTS AND CONCLUSION: The survival rate of rat hepatocytes was 85-95%. At 14 days, the survival rate of transplantation group (75%) was significantly higher than that of control group (30%) (P=0.01). Hepatic function improvement of transplantation group was significantly better than that of control group. At 30 days after transplantation, immunofluorescent staining of albumin could be seen in spleen; at 15 days after transplantation, transplanted hepatocytes were found in spleen. There were many influence factors in the success of hepatocyte separation, such as collagen enzyme, pH value, perfusion fluid and perfusion method. Hepatocyte intraplenic transplantation could improve the survival rate and ameliorate the hepatic function of rats with acute hepatic failure.