The Expression And Significance Of Angiopoietin1, 2 And Tie2 On Vascular Endothelial Cells And In Vitro Model Three-dimensional Vascular-like Structure

Objective We create a three dimension(3D) in vitro model for angiogenesis and detect the expression level of Ang1,2 and Tie2 receptor in human umbilical vein endothelial cells(HUVECs) and three-dimensional vascular-like structure, to explore the effects of Ang1,2 and Tie2 in vitro angiogenesis, and lay a foundation for the further research in the inhibition in vitro angiogenesis in the future.Methods Collagenase was used to isolate the endothelial cell from human umbilical vein, followed by a process of identifying HUVECs by the technique of immunocellochemistry. Three-dimensional vascular-like structure is acquired in vitro three-dimensional culture of endothelial cell in the sandwith culture system. By Semiquantinative Reverse Transcription Polymerase Chain Reaction , we detect the expression level of Ang1,2 and Tie2 mRNA in 10 specimens of in vitro HUVECs, 10 specimens of in vivo HUVECs, and 10 specimens of in vitro three-dimensional vascular-like structure;and detect the expression level of Ang1,2 and Tie2 protein by immunocellochemistry and immunohistochemica method in vitro and in vivo HUVECs.Results1. Single-cell mixing suspension most of which are HUVECs is obtained by filling Collagenase into human umbilical vein. HUVECs are purified by plating cells to the cultivate plating repeatingly. The purified cells are identified to be HUVECs by the technique of immunocellochemistry. Three-dimensional vascular-like structure is acquired in three-dimensional culture system.2. The expression of Ang2 and Tie2 in vitro three-dimensional vascular-like structure are significantly higher than that in vitro and in vivo HUVECs (P<0.05). The expression of Ang2 and Tie2 in vitro HUVECs are not higher than that in vivo HUVECs(P>0.05). 3. The expression level of Ang1 in vitro,in vivo HUVECs and in vitro three-dimensional vascular-like structure have not significant deference(P>0.05).4. the expressions of Ang1,2 and Tie2 protein in vitro HUVECs and in vivo HUVECs are positive, but the expression level has not significant deference(P>0.05).Conclusion1. In vitro three-dimensional cell culture system is a convenient method which has merit in the growth of endothelial cell, the formation of tube-meshworks and the three-dimensional meshwork,and especially suit for the study in the effects of the factors on the angiogenesis.2. Angiopoietin family and receptors take an important part in angiogenesis.