MSCs联合EPCs在组织工程血管化中的作用研究

The Role of MSCs Combined with EPCs in Vascularization of Tissue Engineering

目的:探讨间充质干细胞(MSCs)联合内皮祖细胞(EPCs)体外促进组织工程血管化。方法:利用全骨髓贴壁法结合差时贴壁法分离、扩增小鼠骨髓来源的MSCs和EPCs,运用流式细胞技术测定MSCs和EPCs的表面标记物。取第3代MSCs和EPCs,分别按3×106个接种于无纺布聚乙醇酸支架(PGA)上后体外培养8周(MSCs+EPCs组按1.5×106个以1∶1的比例共同接种)。细胞-支架复合物常规切片行HE染色,显微镜下观察组织形态。通过免疫组化染色评价CD31、CD34在各实验组中的表达情况。结果:组织形态学观察可见MSCs+EPCs组的细胞数目及组织形态较MSCs组、EPCs组多且规则,PGA代谢产物已基本完全降解。免疫组化结果显示CD31、CD34在MSCs组、EPCs组、MSCs+EPCs组中的染色得分为(1.67±0.82)、(1.33±0.52),(4.83±1.33)、(5.33±1.03),(8.67±0.52)、(8.00±2.45),各组差异有统计学意义。结论:MSCs联合EPCs体外可促进组织工程血管化,并且其增殖分化和促血管生成能力高于单一培养组。

Objective：. Study on mesenchymal stem cells （MSCs） combined with endothelial progenitor cells （EPCs） to promote tissue engineering vascularization in vitro.Methods：MSCs and EPCs derived from mouse bone marrow were isolated and expanded by using whole bone marrow adherent method and differential adhesion method,the immune phenotypes of MSCs and EPCs were detected by flow cytometry.Take the third generation of MSCs and EPCs, respectively, by3×106cells inoculated on a non-woven polyglycolic acid （PGA） on the culture in vitro for 8 weeks （MSCs＋EPCs group were inoculated with a ratio of1： 1 in 1.5×106cells）.Cell-scaffold complex sections were stained with HE for histomorphological observation.Immunohistochemical staining to evaluate the expression of CD31,CD34 in each experimental group.Results： Histological observation showed that the number of cells and morphology in MSCs＋EPCs group were better than those of MSCs group and EPCs group, and the metabolites of PGA were al-most degraded.Immunohistochemistry showed that the staining scores of CD31 and CD34 in MSCs group, EPCs group and MSCs＋EPCs group were （1.67±0.82）, （1.33±0.52）, （4.83±1.33）, （5.33±1.03）, （8.67±0.52）, （8.00±2.45）, the dif-ference was statistically significant.Conclusion：MSCs combined with EPCs can promote the vascularization of tissue engineering in vitro, and its proliferation and angiogenesis are higher than that in single culture group.