Primary culture and identification of human placental microvascular endothelial cell in vitro / 第三军医大学学报

ABSTRACT

Objective To explore the methods of culture and identification of human placental microvascular endothelial cells(HPMVEC) in vitro,aiming to construct the polar model of human placental barrier.Methods By proteolytic enzymes,the tissular and cellular suspensions were collected from the chorionic villi of pregnant and centrifugated over a preformed discontinuous Percoll gradient.The highly purified human placental microvascular endothelial cells were obtained and cultured in HG-DMEM with fetal calf serum and identified by immunocytochemistry(SP methods) and transmission electron microscope.Results The highly purified human microvascular endothelial cells were cultured in vitro and passaged for three to five generations.Factor Ⅷ-related antigen and CD34 were both positive by immunocytochemistry.Weibel-Palade body(W-P body) was observed by transmission electron microscope.Conclusion Human placental microvascular endothelial cells can be isolated and cultured successfully in vitro by our method.