大鼠骨髓间充质干细胞体外培养可自发分化为内皮细胞

Spontaneous endothelial cells differentiation from rat bone marrow-derived mesenchymal stem cells in vitro

目的 :探讨大鼠骨髓间充质干细胞(bone marrow-derived mesenchymal stem cells,BMSCs)在体外正常培养条件下能否自发分化为内皮细胞。方法:分离原代大鼠BMSCs,传代后正常培养至第7代,选用第3代和第7代用于本次实验。用Real-time PCR方法检测第3代和第7代BMSCs内皮细胞标识物CD31、血管生成素受体2(tyrosine kinase with immunoglobulin-likeand EGF-like domains 2,Tie-2)及血管性血友病因子(von Willebrand factor,vWF)的表达;体外血管生成试剂盒(Matrigel)检测血管生成能力;免疫荧光染色检测细胞内vWF蛋白的表达。结果:在培养至第7代后,部分BMSCs在形态上呈椭圆形;Real-timePCR和免疫荧光的结果显示第7代BMSCs的内皮细胞标志物表达水平明显高于第3代BMSCs;体外血管生成实验表明第7代BMSCs在Matrigel胶上能形成网状结构。结论:BMSCs在体外正常培养条件下细胞表型会发生一定的变化,并可部分自发分化为内皮细胞。

Objective：To investigate whether rat bone marrow-derived mesenchymal stem cells（BMSCs） could spontaneously differentiate into endothelial cells in normal culture condition.Methods：Primary rat BMSCs were separated and cultured to passage 7.The passage 3 and passage 7 BMSCs were prepared for this experiment.The expression of endothelial cell markers CD31,tyrosine kinase with immunoglobulin-like and EGF-like domains 2（Tie-2） and von Willebrand factor（vWF） in passage 3 and passage 7 BMSCs were determined by Real-time PCR,and the ability of angiogenesis were determined by an in vitro angiogenesis kit（Matrigel）.Futhermore,the expression of vWF protein in BMSCs were detected using immunofluorescence staining.Results：After normal cultured to passage 7,BMSCs partly formed an oval shape.Real-time PCR and immunofluorescence showed that the expression of endothelial cell markers were significantly higher in passage 7 BMSCs than in passage 3 BMSCs.In vitro angiogenesis experiments showed that passage 7 BMSCs could form a mesh structure in matrigel.Conclusion：BMSCs in normal culture condition changed their phenotypes,and could partially differentiate into endothelial cells spontaneously.