Objective To study the isolation,culture and identification of human umbilical vein endothelial cells(HUVECs) in vitro,and analyze antigens expression.Methods Cultured cells were isolated from umbilical cord by enzyme digestion and were cultured in vitro.Cultured cells were subjected to flow cytometer to analyze the expression of CD31,CD34,VWF,KDR and CD133.Immunofluorescence was performed to detect VWF and dual binding of ac-LDL and UEA-1.Results Cultured HUVECs were like slabstones,which were positively stained for CD31,CD34,VWF,KDR,but negative for CD133.Cultured HUVECs could endocytose ac-LDL and UEA-1.Conclusions HUVECs with the characters of endothelial cells can be cultured abundantly by this method.Purity of HUVECs can be assayed by analyzing antigens expression,which can be used to establish the model of endothelial cells.