Optimization purity of isolated primary liver endothelial cells (LSEC) using adenoviral gene transfer

Background: The isolation of murine primary non parenchymal liver cells is crucial for most studies addressing liver pathology. Because of the small organ size of mouse liver and similar cell density (HSC: 1.053 g/ml; Kupffer cells: 1.060 g/ml, liver endothelial cells: 1.080 g/ml) this often results in low yields and poor purity. Unlike hepatocytes and hepatic stellate cells (HSC) liver sinusoidal endothelial cells (LSEC) do not express the integrin receptor alpha(v)beta3 and show low expression of the primary coxsackie-adenovirus receptor (CAR), both of which are required for efficient infection by human adenovirus type 5 (Ad5). Therefore primary LSEC cultures can be further purified by Ad5-mediated gene transfer into contaminating liver cells without compromising LSEC viability.