Experimental Study Of Endothelial Progenitor Cells Transfected With VEGF165 Gene In The Treatment Of Limb Ischemia

Background and ObjectiveWith the change of eating habit,improvement of life and life lengthening, the morbidity of peripheral arterial diseases (PAD) is higher than ever, and lower extremity ischemic diseases are threatening human health seriously. Now the treatments of lower extremity ischemic diseases include medical therapy, surgery and endovascular treatment. But there is always no opportunity of surgery for those who have serious stenosis of ourflow tract especially for those with diabetes, and there is no satisfied treatments yet. Improving the neovascularization and collateral circulation by using some new methods maybe feasible and effective. At present, stem cells therapy and gene therapy are hot spots in the treatments of ischemic diseases. The advantage of stem cells is that it is easier to be modified compared with adult tissues, and it is a good target cell for gene therapy. So combined with stem cell and gene therapy may be promising for the treatment of ischemic diseases. In this experiment, mononuclearcells were isolated from rabbit bone marrow and induced for endothelial progenitor cells(EPCs) in vitro. Then rabbits with ischemic limbs were transplanted with EPCs transfected with VEGF165, the effects of neovascularization were observed. We expect to offer some experimental evidences for the clinical application of stem cell and gene therapy in ischemic diseases.Methods1. Rabbit bone marrow was obtained by iliac crest puncture,then bone marrow mononuclearcells were isolated by gradient centrifugation using Ficoll centrifugate. Mononuclearcells were then cultivated with M199 culture medium and induced to EPCs by VEGF, bFGF and IGF-1. Cultivated cells were identified by immunofluorescence and electron microscope.2. The cultivated cells were transfected with recombined adenovirus, into which...