Selection of in vivo retrovirally transduced hepatocytes leads to efficient and predictable mouse liver repopulation.

Abstract

SPECIFIC AIMSStable liver gene transfer is generally limited by the low efficiency of commonly used vectors. One way to circumvent this difficulty is to confer a selective advantage on transduced hepatocytes, allowing them to progressively repopulate the liver. The aim of this study was to investigate the possibility to repopulate a normal liver to a desired level with in vivo retrovirally engineered hepatocytes.PRINCIPAL FINDINGS1. A retroviral vector encoding Bcl-2 can repopulate the liver up to 85% after 10 wkWe have previously shown that transplanted transgenic hepatocytes expressing Bcl-2 could repopulate a normal mouse liver submitted to repeated Fas apoptotic challenges.We constructed two retroviral plasmids containing either the green fluorescent protein reporter gene alone (MFG-GFP) or a bicistronic cassette including human Bcl-2 cDNA and the GFP reporter gene (MFG-BIG) separated by an internal ribosomal entry site. To compare our liver repopulation approach with direct retroviral transduction, o...