Effects of sera, basic fibroblast growth factor, heparin and cyclic AMP-stimulation on proliferation of human...

The aim of this study was to find culture conditions that optimize the proliferation of human endothelial cells (ECs). The effects of different sera, growth factors and other additives on ECs derived from the adult human great saphenous or the umbilical vein were studied. Human serum (HS) at 10 and 40% was significantly more effective than fetal calf serum at 10 and 30%, respectively. The addition of basic fibroblast growth factor (bFGF) increased proliferation alone and in combination with heparin. Heparin alone increased EC growth only in medium containing 40% HS. The addition of cholera toxin (CT) and a phosphodiesterase inhibitor (IBMX) to raise cAMP-levels, stimulated proliferation of both cell types but was more pronounced on the ECs from the saphenous vein. The cAMP-levels were elevated equally in both cell types. However, db-cAMP stimulated proliferation only of the ECs from the saphenous vein. An additive stimulatory effect was observed when bFGF and CT/IBMX were combined. For saphenous vein ECs, a medium containing HS (40%), bFGF, heparin, CT and IBMX was found optimal for proliferation. We conclude that these compounds may be used to increase EC growth and that, even a limited number of donor ECs may be sufficient starting material for in vitro studies on the human endothelium.