BACKGROUND: The establishment of human umbilical vein endothelial cells model in vitro has significant meaning for the study of neovascularization, but human umbilical vein endothelial cells are difficult to cultivate in vitro and there is not a criterion.
OBJECTIVE: To explore how to harvest and identify human umbilical vein endothelial cells in vitro.
METHODS: 0.25 g/L Trypsin and 0.02% ethylene diamine tetraacetic acid perfusion method was used to isolate human umbilical vein endothelial cells from the fresh umbilical cord, and the cells were cultured and amplified. The composition of culture medium was simplified by not adding vascular endothelial cell growth factor and heparin. Human umbilical vein endothelial cells grown to 80% confluence were identified by morphological observation and immunofluorescence method.
RESULTS AND CONCLUSION: The endothelial cells spread on the bottom of the dishes within 2 hours, then coalesced and grew to form a confluent monolayer of polygonal cells within 24 hours. The cultured cells were identified as human umbilical vein endothelial cells. Trypsin perfusion is a simple and effective method for collection of human umbilical vein endothelial cells. Cells harvested with this protocol can be used as models on research of vascular endothelial cells in vitro.