The proliferation of human umbilical vein endothelial cells in serum-free medium

Experimental conditions have been defined to allow human umbilical vein endothelial cells to grow in the complete absence of serum. Endothelial cells maintained in fibronectin-precoated plastic dishes could be grown in medium RPMI-1640 supplemented with transferrin, insulin, human serum albumin (HSA) and epidermal growth factor (EGF) or fibroblast growth factor (FGF). Endothelial cell growth supplement (ECGS) stimulated the proliferation in the presence of EGF or FGF, but its presence is not absolutely required to obtain growth of endothelial cells in serum-free medium. Serum was only required for a short period to allow attachment and spreading of the cells after trypsinization. Endothelial cells grown to confluence in serum-free medium exhibited morphological characteristics comparable to cells cultured in the presence of human serum, secreted normal quantities of factor VIII-related antigen into the culture medium, and synthesize prostacyclin.