[An effective method of isolation and primary culture of human umbilical arterial endothelial cells].

ABSTRACT

OBJECTIVE: To explore and establish an efficient system for isolation and culture of human umbilical arterial endothelial cells (HUAECs). METHODS: The human umbilical arteries were perfused and washed by PBS, and then digested by 0.1 g/L type 1 collagenase. Digested cell supernatants were collected, centrifuged, and the re-suspended cells were cultured in a specific medium. The identification of endothelial cells was carried out by observing the cellular morphology, immunostaining the endothelial surface marker CD31 under fluorescent microscopy and examining the capability to form tube-like structure. RESULTS: The isolated cells grew in a monolayer and exhibited mosaic-like pattern under an inverted phase-contrast microscope. They strongly expressed CD31, an endothelium-specific surface marker, and had an obvious capability to form tube-like structure. CONCLUSION: The method established for isolation of HUAECs in this experiment is simple, rapid and efficient. In addition, the isolated cells are of high purity and high survival.