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1 Departments of Medicine and 2 Cell Biology, Duke University Medical Center, Durham, North Carolina 27710
Adenovirus-mediated
gene transfer has become an important tool with which to introduce
genetic material into cells. Available data emphasize efficient
adenoviral transduction of parenchymal liver cells (i.e., hepatocytes)
in both in vitro and in vivo model systems, typically in normal cells.
The aim of this study was to evaluate gene transfer to nonparenchymal
(and parenchymal) cells of the normal and injured rat liver.
Hepatocytes, stellate cells, and endothelial cells were isolated by
standard methods. Liver injury was induced by bile duct ligation or
carbon tetrachloride administration. Cells were transduced in vitro
with an adenovirus encoding 
-galactosidase (Ad.-gal) over a range
of viral titers, and transduced cells were identified by detection of
X-gal. In vivo transduction efficiency was studied in normal and
injured livers using cell isolation techniques. Nonparenchymal cells
were transduced with greater frequency than hepatocytes at all
adenoviral titers tested, both in vitro and in vivo. After liver
injury, adenoviral transduction was reduced for all liver cell types
compared with that for cells from normal livers (at all virus titers). Notably, transduction efficiency remained greater in nonparenchymal cells than in hepatocytes after liver injury. This work implies that,
to achieve comparable gene expression in the injured liver, higher
adenoviral titers may be required, an important consideration as gene
therapy in disease states is considered.
cirrhosis; bile duct ligation; carbon tetrachloride; stellate cell; sinusoidal endothelial cell
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