中国组织工程研究

中国组织工程研究 ›› 2011, Vol. 15 ›› Issue (6): 972-975.doi: 10.3969/j.issn.1673-8225.2011.06.006

• 骨髓干细胞 bone marrow stem cells • 上一篇    下一篇

全反式视黄酸诱导兔骨髓间充质干细胞向神经细胞的分化

周文逊1，2，张雁儒1   

  1. 1郑州大学基础医学院，河南省郑州市  450064
    2漯河医学高等专科学校，河南省漯河市  462002
  • 收稿日期:2010-11-02 修回日期:2010-12-14 出版日期:2011-02-05 发布日期:2011-02-05
  • 通讯作者: 张雁儒，硕士，副教授，郑州大学基础医学院人体解剖学教研室，河南省郑州市 450064 zhangyanru818@yahoo.com.cn
  • 作者简介:周文逊★，男，1982年生，湖北省潜江市人，汉族，郑州大学基础医学院在读硕士，讲师，主要从事人体解剖学研究。 zwx0929@163.com

All-trans-retinoic acid induces the differentiation of rabbit bone marrow mesenchymal stem cells into neural cells

Zhou Wen-xun1,2, Zhang Yan-ru1   

  1. 1Basic Medical College, Zhengzhou University, Zhengzhou  450064, Henan Province, China
    2Luohe Medical College, Luohe  462002, Henan Province, China
  • Received:2010-11-02 Revised:2010-12-14 Online:2011-02-05 Published:2011-02-05
  • Contact: Zhang Yan-ru, Master, Associate professor, Basic Medical College, Zhengzhou University, Zhengzhou 450064, Henan Province, China zhangyanru818@yahoo.com.cn
  • About author:Zhou Wen-xun★, Studying for master’s degree, Lecturer, Basic Medical College, Zhengzhou University, Zhengzhou 450064, Henan Province, China; Luohe Medical College, Luohe 462002, Henan Province, China zwx0929@163.com

PDF

451

被引次数

11

摘要：

背景：目前，在骨髓间充质干细胞体外向神经细胞分化的实验中，较多采用抗氧化剂法和细胞因子法，但诱导效率低。
目的：探讨全反式视黄酸在兔骨髓间充质干细胞向神经细胞分化中的作用。
方法：体外分离培养兔骨髓间充质干细胞，取第4代骨髓间充质干细胞观察细胞形态，实验组用0.4 μmol/L全反式视黄酸预诱导24 h后，改用神经细胞培养基继续培养。神经细胞培养基培养4，8，16及24 h，免疫组织化学检测巢蛋白、神经元特异性烯醇化酶和胶质纤维酸性蛋白的表达情况，采用RT-PCR的方法检测巢蛋白和神经元特异性烯醇化酶的表达水平。
结果与结论：骨髓间充质干细胞经培养、传代后，细胞贴壁生长，呈长梭形。免疫组织化学检测结果显示：全反式视黄酸诱导组巢蛋白及神经元特异性烯醇化酶呈阳性，诱导后细胞的活力良好。RT-PCR结果显示全反式视黄酸诱导组巢蛋白在诱导前后均有表达，神经元特异性烯醇化酶在诱导后16 h可见明显的扩增条带，24 h后更加明显。提示全反式视黄酸能够在体外促进兔骨髓间充质干细胞向神经细胞分化。

关键词: 全反式视黄酸, 骨髓间充质干细胞, 诱导, 神经细胞, 分化

Abstract:

BACKGROUND: At present, antioxidant and cytokine method are used in the study of the differentiation of bone marrow mesenchymal stem cells (BMSCs) into nerve cells in vitro, but the induced efficiency was low.
OBJECTIVE: To investigate the role of all-trans-retinoic acid (ATRA) in the differentiation of rabbit BMSCs into neural cells.
METHODS: Rabbits BMSCs were isolated and expanded in vitro, and the 4th generations were obtained for observation of cell morphology. After 24 hours of induction using 0.4 μmol/L ATRA, neuronal medium was used in the experimental group. At 4, 8, 16 and 24 hours following culture in the neuronal medium, immunohistochemistry was used to detect expressions of nestin, neuron specific enolase (NSE) and glial fibrillary acidic protein (GFAP). RT-PCR was utilized to determine expression of nestin and NSE.
RESULTS AND CONCLUSION: Following culture and subculture, BMSCs were adherent and fibroblast-like. Immunohistochemical method demonstrated that ATRA group was positive for nestin and NSE. Following induction, cell viability was good. RT-PCR results showed that ATRA group expressed nestin before and after induction. At 16 hours following induction, NSE exhibited significant amplification band, and became more obvious at 24 hours. These suggest that ATRA could promote the differentiation of rabbit BMSCs into neural cells in vitro.

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