Gene expression in implanted rat hepatocytes following retroviral-mediated gene transfer

An hepatocyte transplantation-gene transfer protocol has been developed whereby liver cells containing an expressing Neo^Rgene can be successfully implanted in vivo. Adult primary cultures of rat hepatocytes, after infection with the retroviral vector N2, were grown on a floating solid support (coated with purified collagen IV) in a serum-free hormonally defined medium designed for hepatocytes that also contained G418. Under these conditions, normal adult hepatocytes expressing the Neo^Rgene could be grown to high density. The solid supports holding the gene-engineered hepatocytes were then implanted into adult rats into subcutaneous and intraperitoneal sites. After one to two weeks, the supports were removed and shown to still contain the gene-engineered hepatocytes expressing the Neo^Rgene. These results suggest that cells from solid organs, such as the liver, are potential targets for gene transfer and expression studies in vivo.