骨髓间充质干细胞体外构建工程化心肌组织的生存

背景：到目前为止，工程化心肌组织仍然面临很多的问题。研究证实，骨髓间充质干细胞可分化为心肌样细胞；聚乙交酯、聚己内酯为常用的人工高分子材料，生物相容性良好。目的：观察体外构建骨髓间充质干细胞-聚乙交酯-聚己内酯共聚物补片在正常心肌组织和梗死后心肌组织中的生长情况。方法：SD大鼠骨髓间充质干细胞采用贴壁分离筛选法进行分离、培养，选取第3代进行体外DAPI标记。制作骨髓间充质干细胞悬液(2×106/cm2)种植于聚乙交酯-聚己内酯共聚物上形成骨髓间充质干细胞-聚乙交酯-聚己内酯共聚物补片。培养48 h在电镜下观察；苏木精-伊红染色后在光学显微镜下观察。大鼠结扎左冠状动脉前降支制作心肌梗死模型。将补片植入到正常心肌组织和梗死后心肌组织培养5周，行病理学检查了解骨髓间充质干细胞在组织中的存活情况。结果与结论：光学显微镜和电镜观察结果示，骨髓间充质干细胞在聚乙交酯-聚己内酯共聚物支架中呈三维生长，细胞与支架黏附良好。在激光共聚焦显微镜下，对比观察补片植入正常心肌组织后第1周和第5周切片结果，同第1周相比，第5周的心肌组织内出现DAPI标记的骨髓间充质干细胞；心肌梗死区出现DAPI标记的骨髓间充质干细胞。苏木精-伊红染色结果示，梗死区出现骨髓间充质干细胞。提示骨髓间充质干细胞在聚乙交酯-聚己内酯共聚物支架上贴附生长，黏附良好。骨髓间充质干细胞-聚乙交酯-聚己内酯共聚物补片可用于心肌组织修复。中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程全文链接：

Survival of engineered myocardium constructed by bone marrow mesenchymal stem cells in vitro

BACKGROUND: So far, engineered myocardium is still facing many problems. Research has demonstrated that bone marrow mesenchymal stem cells can be differentiated into myocardial cells. Polyglycolide and polycaprolactone are commonly used artificial polymers, which have good biocompatibility. OBJECTIVE: To observe the growth of the poly(glycolic acid)/poly(e-caprolactone) copolymer patch in vitro in normal myocardium and infarcted myocardium.METHODS: Bone marrow mesenchymal stem cells in Sprague-Dawley rats were separated using adherent separation and selection method, cultured in vitro. The third passage was labeled with DAPI. The bone marrow mesenchymal stem cell suspensions (2×106/cm2) were produced and planted on poly(glycolic acid)/ poly(e-caprolactone) copolymer scaffolds to form poly(glycolic acid)/poly(e-caprolactone) copolymer patch. After culturing for 48 hours, the specimens were observed under electron microscope, stained with hematoxylin and eosin, and then observed under light microscope. Rat models of myocardial infarction were established by ligating left anterior descending coronary artery. Poly(glycolic acid)/poly(e-caprolactone) copolymer patch was implanted into the normal and infarcted myocardium for 5 weeks. The survival of bone marrow mesenchymal stem cells was determined by the detection of pathology.RESULTS AND CONCLUSION: Results of light microscope and electron microscope demonstrated that bone marrow mesenchymal stem cells grew three-dimensionally on poly(glycolic acid)/poly(e-caprolactone) copolymer patch. Cells and patch were adhesive well. Under laser confocal microscopy, compared with the first week, bone marrow mesenchymal stem cells were marked by DAPI in the myocardium at the fifth week. There were bone marrow mesenchymal stem cells marked by DAPI in the infracted area. Results of hematoxylin-eosin staining exhibited that bone marrow mesenchymal stem cells were detected in the infarct area. These results suggested that bone marrow mesenchymal stem cells adhered to the poly(glycolic acid)/poly(e-caprolactone) copolymer stent well. The complexes of poly(glycolic acid)/poly(e-caprolactone) copolymer and bone marrow mesenchymal stem cells can be used for reparation of myocardium.