Transplantation Of Vascular Endothelial Progenitor Cells With Modificatory VEGF165 Gene In The Treatment Of Chronic Deep Venous Thrombosis

Objective: To study the therapeutic effect of endothelial progenitor cells with modificatory VEGF165 gene on the chronic deep venous thrombosis.Methods:1. The bone marrow mononuclear cells of Wistar rats were isolated by Ficoll and cultured with EBM-2MV medium cultivation. 2. Observe the features of culture in vitro and biological characteristics of EPCs. MTT assay were used to draw growth curve of cells.3. After amplified by human embryo renal cells (HEK293A), the adenoviruses were used to transfect EPCs in suitable titer. Then marked with Dil in vitro.4. Experimental rat models(about 300g) of chronic deep vein thrombosis were obtained by partial ligation of inferior vena cava. The rats were randomly divided into four groups, and 16 animals in each group.5. The cells transplantation is through the femoral vein. A group:Ad-VEGF165 transfection group,1ml 106 Ad-VEGF165-EPCs transplantation;B group:EPCs group,1ml 106 EPCs transplantation; C group:Ad-GFP transfection Group, 1ml 106 Ad-GFP-EPCs transplantation; D group:control group,1 ml of medium transplantation. 6. Harvest the thrombus section vena cava inferior and the thrombus at different time. HE staining and vWF immunohistochemical staining were performed to detect the recanalization of thrombus, and the neovascularization was observed by scanning electron microscopy. The capillary density was determined quantitatively by counting the capillaries under high-power microscope.Results:1 Bone marrow mononuclear cells were isolated by Ficoll and EPCs were induced with EBM-2MV medium . 2 Adenoviruses in high titer were obtained through amplificationin HEK293A and transfected EPCs effectively.3 The method of rat model of chronic deep venous thrombosis was simple and reliable. The effect is accurate.4 Transplantation of vascular endothelial progenitor cells transfected with VEGF165 gene promoted the thrombus organization and revascularization.Conclusion:The transplantation of EPCs transfected with Ad-GFP-VEGF165 promoted the thrombus organization and recanalization significantly.