Establishment of a human placental endothelial cell line with extended life span after transfection with SV 40 T-antigens

Studies utilizing microvascular human endothelial cells are relatively few because of substantial difficulties in isolation and cultivation, respective limited proliferation capability and functional variation of primary cells. To facilitate the closer examination of capillary endothelial characteristics, we isolated microvascular endothelial cells from full-term placenta and transfected the cells via lipofection with pRNS-1, encoding for the small and large T-antigens of SV 40. One cell line, HPEC-A1, with a largely extended life span was isolated and kept in culture for up to 80 cumulative population doublings. The cell line HPEC-A1 expressed SV 40 T-antigens and exhibited endothelial characteristics identical to primary cells, such as the uptake of low density lipoprotein, binding of Ulex europaeus lectin I, and the expression of von Willebrand factor, thrombomodulin-α, VE-cadherin, and a set of integrins.