Effects of platelets and certain platelet components on growth of cultured human endothelial cells.

Abstract

INTRODUCTION The recent successful growth of human and nonhuman endothelial cells in tissue culture in several laboratories 1-6 has opened the way for a study of factors that may influence the rate of replication of these cells. One laboratory has reported that the presence of platelets in the nutrient medium enhances the growth of endothelial cells. 1 We have confirmed this report and have extended these observations to include the effects of certain platelet components on endothelial cell replication. The importance of these studies lies in the fact that platelets and certain of their components might influence the rate at which endothelial cells grow in vivo as well as in vitro. METHODS Growth of Human Endothelial Cells in Tissue Culture. Techniques for recovery of endothelial cells from human umbilical cord vein by use of collagenase have been reported. 2 Isolated endothelial cells were washed once in modified (Ca++ and Mg++ free Tyrode's solution (MT'S)) and twice in medium 199 containing 20% fetal calf serum. Isolated cells were 83 to 96% viable by the erythrosin B dye exclusion test. Cells were isolated from umbilical cord vein under strict sterile conditions. Cell cultures were carried out in polystyrene culture flasks (Falcon Plastics, Los Angeles, Cal.) with a growth area of 25 cm. 2 Culture flasks were seeded with approximately 500,000 cells per flask with the cells at a final:concentration of 500 per mm3.