基础医学与临床 ›› 2016, Vol. 36 ›› Issue (4): 513-519.

• 研究论文 • 上一篇    下一篇

胰岛素样生长因子结合蛋白-4对大鼠骨髓间充质干细胞增殖和神经分化的影响

李慧文1,于树夔2,郝飞2,牛含虚2,徐群渊3,段德义3   

  1. 1. 首都医科大学基础医学院
    2. 首都医科大学 基础医学院 神经生物学系
    3. 首都医科大学北京神经科学研究所
  • 收稿日期:2015-11-26 修回日期:2016-01-12 出版日期:2016-04-05 发布日期:2016-03-29
  • 通讯作者: 段德义 E-mail:duandy@ccmu.edu.cn

Impacts of IGFBP4 on proliferation and neural differentiation of bone marrow mesenchymal stem cells of rats

  • Received:2015-11-26 Revised:2016-01-12 Online:2016-04-05 Published:2016-03-29

摘要: 目的 探究胰岛素样生长因子结合蛋白4(IGFBP4)对骨髓间充质干细胞(BMSC)增殖和神经分化的影响。方法 通过全骨髓贴壁法分离培养BMSCs,免疫荧光检测IGFBP4在P1、P4、P6、P8和P10 BMSCs的表达。取第4代BMSC细胞,分为单纯培养基组和添加IGFBP4组,用CCK8法检测吸光光度值。再将BMSC细胞分为3组,诱导培养基NeuroCult培养再分别添加EGF+bFGF和EGF+bFGF+IGFBP4,诱导其向神经祖细胞分化,免疫荧光染色检测Nestin和Sox-2表达,并观察神经球数量和细胞增殖活性。结果 成功获得原代大鼠BMSCs,可见CD105、CD44和CD90表达,IGFBP4的表达随培养代数而增加,外源IGFBP4能明显抑制BMSCs的增殖(P<0.05)。经诱导BMSCs形成的神经球样结构表达Nestin和Sox-2,细胞增殖活性升高(P<0.05)。结论 IGFBP4能抑制BMSCs的增殖,促进其向神经祖细胞的分化。

关键词: 胰岛素样生长因子结合蛋白-4, 骨髓间充质干细胞, 神经分化

Abstract: Objective To explore the effects of insulin-like growth factor binding protein-4 (IGFBP4) on the proliferation and neural differentiation of bone marrow mesenchymal stem cell (BMSC). Methods BMSCs from SD male rats were isolated by the Whole Marrow Adherence Method and identified using immunofluorescence. IGFBP4 expression in the P1, P4, P6, P8 and P10 BMSCs was detected. P4 BMSCs were divided into two groups, and their proliferation activity was detected by CCK8 assay after treatment with or without exogenous IGFBP4. The P4 BMSCs were further divided into three groups to evaluate their potentials of neural differentiation after treatment with NeuroCult medium plus EGF+bFGF or EGF+bFGF+IGFBP4. Neural progenitor cells with neurosphere-like structures were identified by immunostaining of Nestin and Sox-2. The number of the induced neurosphere-like structure was counted and the proliferation of neural progenitor cells was assayed. Results BMSCs from SD rats were successfully cultured, and the cells were positive for CD44, CD90 and CD105. The intensity of IGFBP4 expression was increased after passaging, and IGFBP4 could significantly inhibit BMSCproliferation (P<0.05). BMSCs could be induced to form neural progenitor cells which were immunopositive for Nestin and Sox-2. IGFBP4 promoted the differentiation of BMSCs into neural progenitor stem cells, as manifested by an increased size of neurospheres and increased activity of proliferation (P<0.05). Conclusions IGFBP4 could significantly inhibit the proliferation of BMSCs and promote the differentiation of BMSCs into neural progenitor stem cells.

Key words: insulin-like growth factor binding protein-4 (IGFBP4), bone marrow mesenchymal stem cell(BMSC), neural differentiation

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