中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2013, Vol. 8 ›› Issue (22): 2078-2085.doi: 10.3969/j.issn.1673-5374.2013.22.007

• 原著:神经损伤修复保护与再生 • 上一篇    下一篇

非黏附骨髓细胞来源间充质干细胞向神经元样细胞的分化

  

  • 收稿日期:2013-04-28 修回日期:2013-06-30 出版日期:2013-08-05 发布日期:2013-08-05

Neuronal-like cell differentiation of non-adherent bone marrow cell-derived mesenchymal stem cells

Yuxin Wu1, Jinghan Zhang2, Xiaoming Ben3   

  1. 1 Department of Neurosurgery, Nanjing Children’s Hospital of Nanjing Medical University, Nanjing 210008, Jiangsu Province, China
    2 Department of Newborn Medicine, Nanjing Children’s Hospital of Nanjing Medical University, Nanjing 210008, Jiangsu Province, China
    3 Department of Newborn Medicine, Shanghai First Maternity and Infant Hospital, Tongji University School of Medicine, Shanghai 200040, China
  • Received:2013-04-28 Revised:2013-06-30 Online:2013-08-05 Published:2013-08-05
  • Contact: Xiaoming Ben, Ph.D., Professor, Department of Newborn Medicine, Shanghai First Maternity and Infant Hospital, Tongji University School of Medicine, Shanghai 200040, China, benxm@163.com.
  • About author:Yuxin Wu, Associate chief physician. Yuxin Wu and Jinghan Zhang contributed equally to this work.
  • Supported by:

    国家自然科学基金(30471836)

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摘要：

实验采用Pour-off法培养C57BL/6J小鼠非黏附骨髓细胞来源间充质干细胞，发现其在体外可形成成纤维样细胞集落形成单位，且内皮细胞生长因子对成纤维样细胞集落形成单位的形成有促进作用。免疫细胞化学染色显示，经碱性成纤维细胞生长因子/内皮细胞生长因子/神经生长因子诱导的非黏附骨髓细胞来源间充质干细胞表达神经元特异性标记物神经丝200和NeuN。取β-galactosidase转基因小鼠的非黏附骨髓细胞来源间充质干细胞，将其移植到脑缺血C57BL/6J小鼠右侧纹状体，8周后，缺血组织可见LacZ染色及抗β-galactosidase免疫组化染色阳性细胞，同时免疫组化双标染色可见?-galactosidase和NeuN的共表达。可见小鼠非黏附骨髓细胞来源间充质干细胞在体内外均可向神经元样细胞分化。

关键词: 神经再生, 干细胞, 非黏附骨髓间充质干细胞, 神经元样细胞, 成纤维样细胞集落形成单位, 增殖, 分化, β-Gal转基因小鼠, 细胞移植, 脑缺血, 骨髓间充质干细胞, 基金资助文章, 图片文章

Abstract:

Non-adherent bone marrow cell-derived mesenchymal stem cells from C57BL/6J mice were sepa-rated and cultured using the “pour-off” method. Non-adherent bone marrow cell-derived mesenchymal stem cells developed colony-forming unit-fibroblasts, and could be expanded by supplementation with epidermal growth factor. Immunocytochemistry showed that the non-adherent bone marrow cell-derived mesenchymal stem cells exposed to basic fibroblast growth fac-tor/epidermal growth factor/nerve growth factor expressed the neuron specific markers, neurofilament-200 and NeuN, in vitro. Non-adherent bone marrow cell-derived mesenchymal stem cells from β-galactosidase transgenic mice were also transplanted into focal ischemic brain (right corpus striatum) of C57BL/6J mice. At 8 weeks, cells positive for LacZ and β-galactosidase staining were observed in the ischemic tissues, and cells co-labeled with both β-galactosidase and NeuN were seen by double immunohistochemical staining. These findings suggest that the non-adherent bone marrow cell-derived mesenchymal stem cells could differentiate into neuronal-like cells in vitro and in vivo.

Key words: neural regeneration, stem cells, non-adherent bone marrow cell-derived mesenchymal stem cells, neuronal-like cells, colony-forming unit-fibroblasts, proliferation, differentiation, beta-galactosidase transgenic mouse, cell transplantation, cerebral ischemia, bone marrow cells-derived mesenchymal stem cells, grants-supported paper, neuroregeneration