骨髓间充质干细胞和内皮祖细胞移植促进血流重建的比较

目的:比较骨髓间充质细胞(Bone Marrow Mesenchymal Stem Cells,BM/MSC)和骨髓源内皮祖细胞(Bone Marrow Endothelialprogenitor cells,BM/EPC)移植促进血流重建的效果,为进一步优化骨髓干细胞移植治疗肢体缺血提供理论基础。方法:获取Lewis大鼠骨髓单个核细胞,在体外培养分化为MSC和EPC。采用Lewis大鼠建立单侧后肢缺血模型。在模型建立后3天,将0.8mlD-Hanks液注入大鼠缺血侧后肢,为对照组(n=6);将8×106个骨髓MSC植入大鼠缺血侧后肢,为MSC组(n=6);将体外培养的8×106个EPC植入大鼠缺血侧后肢,为EPC组(n=6)。细胞移植后3周行缺血大鼠后肢动脉造影,检测缺血侧后肢侧支血管数;获取缺血侧后肢腓肠肌,分别行CD31和α-SMA免疫组化染色,计算毛细血管密度和小动脉密度。结果:MSC组与EPC组侧支血管数无显著性差异,二者均高于对照组;EPC组毛细血管密度明显高于MSC组,二者均高于对照组;MSC组与EPC组小动脉密度无显著性差异,二者均高于对照组。结论:骨髓间充质干细胞移植和内皮祖细胞移植均能够明显促进血流重建,而且骨髓间充质干细胞在治疗肢体缺血性疾病中的优势应该受到重视。

The Comparison of Revascularization after Bone Morrow Mesenchymal StemCells and Endothelial Progenitor Cells Transplantation

Objective: To compared the revascularization after bone marrow mesenchymal stem cells(BM/MSC) and bone marrow endothelial progenitor cells(BM/EPC) transplantation and investigate the roles of different stem cells in revascularization.Methods: MNC were harvested from Lewis rats bone marrow.The MSC and EPC were obtained after MNC were cultured for transplantation.Lower limb ischemic model was made with Lewis rats.After limb ischemic model were made,0.8ml D-Hanks,8×106 MSCs and 8×106 EPCs were injected into control group(n=6),MSC group(n=6) and EPC group(n=6) respectively.At 3 weeks after D-hanks injection/cells transplantation,the quantity of collateral was observed with digital subtraction angiography(DSA).Meanwhile,the CD31and α-SMA of the gastrocnemius muscle obtained from the ischemic limb were tested with immunohistochemical stain.Results: Compared with MSC group,the density of the capillaries in EPC group was significantly higher,and there were significant differences between control group and treatment groups.There was no significant difference between MSC group and EPC group in the number of collaterals and density of arterioles.Conclusion: Both MSC and EPC transplantation can improve revascularization significantly.The roles of revascularization of MSC should be attached importance to in the stem cells transplantation to treat for limb ischemic disease.