大鼠骨髓间充质干细胞体外培养可自发分化为内皮细胞
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国家自然科学基金资助 (30973534,81173052)


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

    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-like and 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-time PCR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。

    Abstract:

    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.

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王露露,胡 亮,刘 超,周 露,李庆平.大鼠骨髓间充质干细胞体外培养可自发分化为内皮细胞[J].南京医科大学学报(自然科学版),2013,(8):1039-1043

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  • 收稿日期:2013-04-17
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  • 在线发布日期: 2013-07-30
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