EXPRESSION OF SV40 TUMOR-ANTIGENS ENABLES HUMAN ENDOTHELIAL-CELLS TO GROW INDEPENDENTLY FROM FETAL CALF SERUM AND EXOGENOUS GROWTH-FACTORS

Human endothelial cells were transfected with a plasmid containing the coding sequence of the large T protein of simian virus 40. Transfected cells were selected for their ability to grow in defined medium (DM). Several cell lines were derived and characterized in their response to endothelial cell growth supplement (ECGS), epidermal growth factor (EGF) and insulin (INS). In addition to cell lines that were dependent on these additives, others growing without any exogenous growth factor could be selected. No evidence of autocrine growth stimulation was found. For growth studies, a simple assay was used based on the acid phosphatase activity as a parameter for the cell number. Cell lines in defined medium showed less chromosome aberrations than those grown in serum-containing medium. Because of their long in vitro life span of about 100 generation doublings and defined medium requirements these cells represent valuable test material for all kinds of investigations on the vascular endothelium.