体外诱导大鼠骨髓间充质干细胞向血管内皮细胞分化的研究

目的: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,体外扩增,流式细胞仪检测MSCs表面抗原表达,以含VEGF、bFGF的诱导分化培养液定向诱导传代使细胞向血管内皮细胞分化,流式细胞仪进行内皮细胞特异性标志物鉴定,测定内皮细胞NO的释放量,电镜观测W-P小体.结果:MSCs在体外传代扩增后流式细胞仪检测结果显示CD29表达阳性,CD34、CD45为阴性,分化后的细胞具有内皮细胞的形态学特征并具特异性W-P小体,可表达CD34,而不表达CD45,能释放NO.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.

In Vitro Differentiation into Endothelial Progenitor Cells from Rat Bone Marrow Mesenchymal Progenitor Cells

Objective To investigate the model of separation and culture and the capability of differentiation into endothelial progenitor cells of bone marrow mesenchymal cells (MSCs) Methods MSCs were separated from bone marrow with density gradient centrifugation,wall sticking screening and amplified in vitro. The surface antigens of MSCs were evaluated with FACS (fluorecene activated cell sorter). The passage cells were induced to differentiate into endothelial cells by means of differentiation induation media containing VEGF and bFGF. The differentiated cells were detected by specific marker by FACS,release of oxide(NO) from them was also detected by NO detection regent and W-P (Weibel-Palade) bodies in the cytoplasm were observed using transmission electron microscopy. Results After being propagated and amplified in vitro,MSCs were positively expressed for CD29 and negatively expressed for CD34 and CD45. The differentiated cells demonstrated the character of endothelial cells under phase contrast microscopy. W-P bodies were found in the induced cells. The CD34 expression levels of the cells,in the same time,NO content in the culture media was detected. Conclusion VEGF and bFGF can induce bone marrow mesenchymal cell into endothelial cells in vitro.