间充质干细胞定向诱导为内皮细胞的体外研究

An Experimental Study on the Differentiation of Bone Marrow Mesenchymal Stem Cells into Endothelial Cells in vitro

目的:为寻找心血管组织工程新的种子细胞来源,探讨大鼠骨髓间充质干细胞(MSCs)的体外培养方法和向血管内皮细胞分化的能力。方法:利用淋巴细胞分离液分离出MSCs,并体外扩增,激光共聚焦显微镜检测表面抗原表达,以含血管内皮细胞生长因子(VEGF)、碱性成纤维生长因子(bFGF)的诱导分化培养液定向诱导传代使其向血管内皮细胞方向分化,观察细胞形态的变化,检测内皮细胞特异性标志物的表达。结果:MSCs在体外传代扩增后激光共聚焦显微镜检测结果显示CD44、CD90表达阳性,CD31、CD45为阴性,分化后细胞具有内皮细胞的形态学特征,并表达内皮细胞特异性标志物CD31。结论:骨髓间充质干细胞可在体外诱导向内皮细胞方向分化。

Objective： To explore methods of separating and culturing mesenchymal stem cells （MSCs） of rats in vitro and to study the differentiation capability of bone marrow MSCs into endothelial cells, so as to provide new source of seed ceils for tissue engineering. Methods： MSCs were separated from bone marrow with density gradient centrifugation and wall sticking screening, and amplified in vitro. The surface antigens of MSCs were evaluated with confocal laser scanning microscope （CLSM）. The passage ceils were induced by differentiation induation media containing VEGF and bFGF to differentiate into endothelial cells. The morphological changes of differentiated cells were observed, and their specific indicator of endothelial ceils was detected. Results： After MSCs being propagated and amplified in vitro, CD44 and CD90 were positively expressed in them, while expression of CD31 or CD45 was negative. The differentiated cells demonstrated the character of endothelial ceils under phase contrast microscope. The expression of endothelial cell specific indicator, CD31, was detected at the same time. Conclusion： Bone marrow mesenchymal stem cells can be induced into endothelial cell line in vitro.