Microvascular endothelial cell culture

Abstract

Microvascular endothelial cells play a central role in inflammation, tumor metastasis, and wound healing. Methods to study these processes in vitro using cells isolated from adult skin, from the inner and the outer segments of the neontal foreskin, and from experimental animals are reviewed. A new modified Iscove's medium supplemented with 2% pre-partum maternal serum, dibutyryl cyclic AMP, isobutyl methylxanthine, thymidine, and hypoxanthine is described. This modified medium supports growth of both adult and neonatal endothelial cells up to seven passages with retention of cytologic markers closely identified with endothelial cells (Weibel-Palade bodies, Factor VIII-associated antigen). Several functions associated with the microvasculature in situ are expressed by microvascular endothelial cells in cell culture. Such functions include the formation of a basement membrane, angiogenesis, intercelluar gap formation in response to vasoactive agents, and the attachment and migration of lymphocytes through endothelial monolayers.