Research on gene transfer of hepatocyte growth factor in hepato-spleno-surgery

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We manufactured adenovirus vector containing hepatocyte growth factor (HGF), of which cDNA was generously provided by Dr.T.Nakamura (Biomedical Research Center, Osaka University School of Medicine), according to the protocol Dr.I Saito (Institute of Medical Science, University of Tokyo) invented.After adenovirus vector containing HGF (AxHGF) was transfected into Hela cells. we collected cultured media which contained 1000-3000 ng/ml of HGF.Bioactivity of HGF produced by AxHGF-transfected Hela cells were proved by proliferative effects on cultured vascular endothelial cells and protective effects on radical-induced hepatocyte injury.AxHGF was propagated in 293 cellsand concentrated with purification by sequential ultracentrifugation in cesium chloride step gradients. In vivo injection of concentrated AxHGF improved histological changes and serum levels of AST, ALT and LDH in the dimethylnitrosoamine-induced cirrhotic rats. Hepatic ischemia-reperfusion injury was also reduced in the AxHGF-transfected rats.Increased angiogenesis could not be observed in the splenic autotransplantation with AxHGF injection. Methods of the experiment and evaluation needed to be reconsidered.Another study utilyzing insulin gene adenovirus vector (AxIns) which we made, demonstrated that AxIns injection promoted liver regeneration and increased body weight after hepatectomy in diabetic rats. One shot injection of AxIns prolonged mean survival periods for 30 days in total pancreatectomized dogs without daily insulin, and FK506, an immunosuppressant, increased the amounts and periods of insulin gene expression by AxIns and prolonged survival periods in AxIns-transfected total pancreatectomized dogs.