Treatment Of Lower Limbs Ischemia Of Experimental Rats By Transplantaion With Endothelial Progenitor Cells From Bone Marrow

Objective: To observe the treatment possibility and effects on rats models withexperimental femoral artery obliterans by transplantaion with endothelial progenitorcells from bone marrow.Methods: Mesenchymal stem cells(MSCs) were separated from bone marrowof male SD rats, with density gradient centrifugation, adherent culture andamplified in DMEM in vitro. Experimental group MSCs were cultured inL-DMEM medium supplemented with VEGF and bFGF.Culture cells were identifiedby immunofluorescenc and immunocytochemistry to analyze the expression ofCD34. Experimental femoral artery obliterans rats models were successfully made.Two weeks later, obtained CD34~+ cells were transplanted into the right lower limbsby i.m.as Grupe A in the 16 rats,and rhe same volume of normal saline into the leftlower limbs as Groupe B as contral. On the 2th week and 4th week, 8 rats weresacrificed respectively and muscles of lower limbs were extracted forimmunohistochemical examinations by CD34. Observe the neoformative bloodcapillary density of the two groups under microscope and count CD34~+ bloodcapillaries respectively, Statistical treatment to analyze the data.Results: Pure MSCs were obtained by density gradient centrifugation andadherent culture. We can directional transform the MSCs into EPCs under acertain condition in vitro. The cultured cells with stem cell characteristics and showedactive proliferative capacity. After the cells transplantation 2 weeks and 4 weeks,ratsin the two groupes were found blood capillary density increased. The average numberof CD34~+ blood capillaries each visual field in Group A is (4.68±1.50) after 2weeks,and Group B is (3.35±1.51) at the same time. After 4 weeks Group A is(6.83±1.78) and Group B is (4.18±1.26).The average number of CD34~- bloodcapillaries each visual field in Group A was more obviously than that in Group B(P＜0.01).Conclusion: We can obtaine EPCs from MSCs in vitro. Transplantation of EPCs byi.m. can incorporate into the local capillary networks and participate vascularizationto improve blood supply of ischemic lower limbs in rats.