Objective To explore the mechanism of mesenchymal stem cells (MSCs) transplantation for chronic hindlimb ischemia in Lewis rats by using cell tracer technique.
Methods MSCs were isolated and cultured by using density gradient centrifugation and adherence method respectively, then labeled by 5-bromo-2-deoxyuridine (BrdU). Eight chronic hindlimb ischemia models of Lewis rats were prepared by using suture-occluded method and then divided randomly to MSCs transplantation group and control group, each group enrolled 4 rats, accepting MSCs transplantation and saline respectively. Then on 7 days and 14 days after transplantation, clinical observation, determination of blood flow, and angiography were performed on rats of the 2 groups. At the same time points after previous tests, rats of the 2 groups were sacrificed to get quadriceps tissues and gastrocnemius tissues to perform HE staining and BrdU immunohis-tochemistry.
Results The 8 rats were all survived on 14 days after transplantation, with no tumor happened and necroses in the transplanted area. On 14 days after transplantation, the blood flow ratio of operated side to un-operated side in the hindlimb (1.773 vs. 1.279) of rats in MSCs transplantation group and control group increased, and the angiography results showed that there were no much increase in ratio of collateral vessels number (0.908 vs. 0.835). There were no significant change in the quadriceps tissues and gastrocnemius tissues by HE staining. The BrdU positive kernels located in the inter-stitial substance cells and vascular endothelia cells, and divided differently in different parts of hindlimb at different time points, that the ratio of positive cells in gastrocnemius tissue was higher than those of quadriceps tissue on 7 days after transplantation, but lower on 14 days.
Conclusions MSCs transplantation can increases the blood perfusion of hindlimb in the early stage of chronic hindlimb ischemia model, and the possible mechanism may be the paracrine effect of MSCs but not the number increase of collateral vessels.
Citation: YANG Shengjia,CHEN Bing,TONG Zhu,ZHANG Shuwen,SHI Xiaolin,XU Qing,ZHANG Bin,ZHANG Jian.. Mechanism of Mesenchymal Stem Cells Transplantation for Chronic Hindlimb Ischemia in Lewis Rat. CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY, 2013, 20(6): 608-614. doi: Copy