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 NeoR gene 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 NeoR gene 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 NeoR gene. These results suggest that cells from solid organs, such as the liver, are potential targets for gene transfer and expression studies in vivo.