Culture and identify the human umbilical vein endothelial cells and investigate the expression of tyrosine kinase-2 with immunoglobulin-like and epidermal growth factor homology domains in the cells

OBJECTIVE: To study the cultural method and identification of human umbilical vein endothelial cells (HUVECs), and investigate the expression of tyrosine kinase-2 with immunoglobulin-like and epidermal growth factor homology domains(Tie-2) in HUVECs. METHODS: HUVECs were isolated from umbilical veins by the technique of irrigative digestion, and were cultivated in plates. The cells were identified by VIII monoclonal antibody. Tie-2 mRNA and protein were detected by reverse transcription-polymerase chain reaction (RT-PCR) and SABC immunocytochemistry. RESULTS: HUVECs could adhere to the plates completely after 24 hours, and confluence a monolayer 4-5 days later. The band of Tie-2 mRNA was obviously and the expression of Tie-2 protein was strongly positive by immunocytochemistry in HUVECs. The positive rate was over 85%. CONCLUSION: Highly purified endothelial cells were isolated. And there were overexpression of Tie-2 in HUVECs.