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 (23): 2171-2177.doi: 10.3969/j.issn.1673-5374.2013.23.007

• 原著:神经损伤修复保护与再生 • 上一篇    下一篇

Noggin和碱性成纤维细胞生长因子的神经诱导分化能力

  

  • 收稿日期:2013-03-15 修回日期:2013-07-08 出版日期:2013-08-15 发布日期:2013-08-15

Noggin versus basic fibroblast growth factor on the differentiation of human embryonic stem cells

Yan Zhang1, Junmei Zhou2, Zhenfu Fang1, Manxi Jiang1, Xuejin Chen1   

  1. 1 Department of Laboratory Animal Sciences, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China
    2 Tissue Engineering Laboratory, Shanghai Institute for Pediatric Research, Xinhua Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200092, China
  • Received:2013-03-15 Revised:2013-07-08 Online:2013-08-15 Published:2013-08-15
  • Contact: Xuejin Chen, Ph.D., Professor, Department of Laboratory Animal Sciences, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China, chenxuej@yahoo.com.cn.
  • About author:Yan Zhang, Master.
  • Supported by:

    上海市基础研究重点项目(No.08JC1413900)

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摘要：

以往研究未见有比较Noggin和碱性成纤维细胞生长因子对人类胚胎干细胞诱导分化为神经前体细胞影响差异的报道。实验通过在无血清的神经诱导培养基中加入100 µg/L Noggin 或20 µg/L碱性成纤维细胞生长因子单层分化诱导作者所在实验室最新建立的人胚胎干细胞H14细胞系分化，诱导2周后，相差显微镜观察发现Noggin诱导细胞中神经花环数量明显大于碱性成纤维细胞生长因子诱导；免疫荧光染色检测结果显示2种诱导方法诱导细胞中大量表达神经前体细胞特征蛋白巢蛋白、β-III微管蛋白和Sox-1；反转录PCR检测显示2种诱导方法诱导细胞中还表达神经前体细胞特征巢蛋白、Sox-1和神经丝蛋白 mRNA，且Noggin诱导的细胞中具有神经前体细胞特征的蛋白和mRNA的细胞数量明显多于碱性成纤维细胞生长因子诱导组。提示在单层分化法平台上，Noggin可能比碱性成纤维细胞生长因子能更快更多的将人类胚胎干细胞诱导分化成神经前体细胞。

关键词: 神经再生, 干细胞, 碱性成纤维细胞生长因子, Noggin, 人胚胎干细胞, 神经前体细胞, 神经分化, 基金资助文章

Abstract:

The difference between Noggin and basic fibroblast growth factor for the neural precursor differen-tiation from human embryonic stem cells has not been studied. In this study, 100 μg/L Noggin or 20 μg/L basic fibroblast growth factor in serum-free neural induction medium was used to differen-tiate human embryonic stem cells H14 into neural precursors using monolayer differentiation. Two weeks after induction, significantly higher numbers of neural rosettes formed in the Noggin-induced group than the basic fibroblast growth factor-induced group, as detected by phase contrast micro-scope. Immunofluorescence staining revealed expression levels of Nestin, β-III Tubulin and Sox-1 were higher in the induced cells and reverse-transcription PCR showed induced cells expressed Nestin, Sox-1 and Neurofilament mRNA. Protein and mRNA expression in the Noggin-induced group was increased compared with the basic fibroblast growth factor-induced group. Noggin has a greater effect than basic fibroblast growth factor on the induction of human embryonic stem cell differentiation into neural precursors by monolayer differentiation, as Noggin accelerates and in-creases the differentiation of neural precursors.

Key words: neural regeneration, stem cells, basic fibroblast growth factor, Noggin, human embryonic stem cells, neural precursors, neural differentiation, grants-supported paper, neuroregeneration