Investigation On The Differentiation And Implantation Of Mesenchymal Stem Cells In Lewis Rat Limb Chronic Ischemia Model

Objective: Establish the animal model (Lewis Rat) of hindlimb ischemia and isolate mesenchymal stem cells, clear the metamorphosing process of stem cells grafts after injecting BrdU labeled MSCs to discuss the exact mechanism in treating arteria ischemic diseases in using stem cells.Method: Surgery fixing, excision of femoral artery along with braches, combinied with endothelial membrane damage and local embolism were adapted in building animal model of hindlimb ischemia. The model was confirmed authentic with laser Doppler analyze of blood flow and blood pressure,histological examination, VIII factor immune histochmistry and angiography of hindlimb; isolated and purified MSCs of rat's bone marrow with gradient centrifugation and adherence to the cultureplastic, then the cells were expanded by subculture successively. The growth curves were drawn, and them orphology was observed In an attempt to analyze immuno and adhesive-phenotype and differentiation properties, the MSCs were evaluated with cytochemical and immunocytochemical methods and labeled with BrdU. Transplant MSCs to the model limb as the offside one injected with physiological saline.Results: After the operation the increased number of small collateral vessels, obvious descent of blood pressure and blood flow were revealed by angiography. adherence to the cultureplastic is one feasible method in culturing MSCs with simple procedure and a high successive rate and it could become a standard way of culturation MSCs. The blood vessels of the transplanted region were proliferated as the function of the limb improved due to one part of the transplanted MSCs formed angioid tissue.Conclusion Mesenchymal stem cells (MSCs) are a multi-potent population which mainly localized in the bone marrow. MSCs were also isolated from the umbilicalblood, peripheralblood, fatty tissue, skin and so on. The special immunity and persist in the xenogeneic explantation of MSCs expand the clinical application. At present the system of culturation, induction and identification of MSCs are gradually becoming complete. MSCs could differentiate into cells derived from mesoderm in the theory meanwhile endothelial cells derived from the mesoderm, so MSCs have potency of differentiated into endothelial cells(ECs). After transplantion of MSCs ,the blood supply of ischemia limb had improved,and the ability of ischemia counteraction was improved.