Immortalization of human endothelial cells by origin-defective simian virus 40 DNA

Human endothelial cells isolated from an umbilical cord vein were transfected with origin-defective simian virus 40 (SV40) DNA. Among several of the SV40 transfected clones isolated, cell lines SV-2 and SV-3 showed a normal endothelial cell morphology and extended life span, and could survive almost 100 generations. Just before crisis, the morphology of SV-3 changed. SV-3T cell line was isolated from this SV-3 culture, which acquired an almost infinite life span, rapid growth rate and the ability to grow in soft agar. At the same time, the SV-3T cell line lost the factor VIII-related antigen and normal endothelial cell morphology, and showed an abnormal chromosome number. Further characterization showed the ability of SV-2 and SV-3T to produce increasing amounts of tissue plasminogen activator and a similar level of a plasminogen activator inhibitor compared with normal human endothelial cells. These results indicate that the SV-3T cell line was transformed and acquired an infinite life span while still r...