中国组织工程研究

中国组织工程研究 ›› 2011, Vol. 15 ›› Issue (27): 4979-4983.doi: 10.3969/j.issn.1673-8225.2011.27.009

• 脂肪干细胞 adipose-derived stem cells • 上一篇    下一篇

多因子联合诱导脂肪组织源性干细胞分化为神经元样细胞

张丽华1，2，汤银娟2，3，董为人2，郭家松2，陈小艳1，王海红2，陈盼盼2，叶泉英1，冯淑兰1，张  易2   

  1. 1佛山科学技术学院医学院组胚教研室，广东省佛山市  528000
    2南方医科大学基础医学院组织学与胚胎学教研室，广东省广州市  510515
    3湘南学院组胚教研室，湖南省郴州市  418000
  • 收稿日期:2011-02-25 修回日期:2011-05-27 出版日期:2011-07-02 发布日期:2011-07-02
  • 通讯作者: 董为人，博士，教授，南方医科大学基础医学院组织学与胚胎学教研室，广东省广州市 510515 wrdong@163.com
  • 作者简介:张丽华★，女，1976年生，广东省五华县人，汉族，2009年南方医科大学毕业，硕士，实验师，主要从事干细胞与组织工程方面的研究。 zlhfs@163.com 并列第一作者：汤银娟★，女，1980年生，湖南省湘潭市人，汉族，2009年南方医科大学毕业，硕士，讲师，主要从事干细胞和组织工程的研究。 Vigor1980@126.com
  • 基金资助:

    佛山市卫生局医学科研立项课题(2010177)；佛山市科技局医学类科技攻关项目(201008063)；广东省高校优秀中青年人才基金资助(C1010086)；广东省自然科学基金资助(8151051501000005)；佛山科学技术学院校级科研课题(2009)；湘南学院一般科研项目(2009Y029)。

Multiple factors-induced differentiation of rat adipose tissue-derived stem cells into neuron-like cells

Zhang Li-hua1, 2, Tang Yin-juan2, 3, Dong Wei-ren2, Guo Jia-song2, Chen Xiao-yan1, Wang Hai-hong2, Chen Pan-pan2, Ye Quan-ying1, Feng Shu-lan1, Zhang Yi2   

  1. 1Department of Histology and Embryology, Medical College of Foshan University, Foshan 528000, Guangdong Province, China
    2Department of Histology and Embryology, Basic Medical College of Southern Medical University, Guangzhou  510515, Guangdong Province, China
    3Department of Histology and Embryology, Xiangnan University, Chenzhou  418000, Hunan Province, China
  • Received:2011-02-25 Revised:2011-05-27 Online:2011-07-02 Published:2011-07-02
  • Contact: Dong Wei-ren, Doctor, Professor, Department of Histology and Embryology, Basic Medical College of Southern Medical University, Guangzhou 510515, Guangdong Province, China wrdong@163.com
  • About author:Zhang Li-hua★, Master, Experimentalist, Department of Histology and Embryology, Medical College of Foshan University, Foshan 528000, Guangdong Province, China; Department of Histology and Embryology, Basic Medical College of Southern Medical University, Guangzhou 510515, Guangdong Province, China zlhfs@163.com Tang Yin-juan, Master, Lecturer, Department of Histology and Embryology, Basic Medical College of Southern Medical University, Guangzhou 510515, Guangdong Province, China; Department of Histology and Embryology, Xiangnan University, Chenzhou 418000, Hunan Province, China Vigor1980@126.com Zhang Li-hua and Tang Yin-juan equally contributed to this paper.
  • Supported by:

    the Scientific Research Program of Foshan Health Bureau, No. 2010177*; the Science and Technology Development Program of Medicine, Foshan Science and Technology Bureau, No. 201008063*; the Young and Middle-aged Talent of Guangdong High Institute, No. C1010086*; the Natural Science Foundation of Guangdong Province, No. 8151051501000 005*; the Scientific Research Program of Foshan University, No. 2009*; General Scientific Research Program of Xiangnan University, No. 2009Y029*

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摘要：

背景：国内均可见有探讨脂肪干细胞向神经元诱导分化的相关研究报道，但运用差速贴壁法纯化脂肪组织源性干细胞，多因子联合诱导脂肪组织源性干细胞分化为神经元样细胞的研究鲜有报道。
目的：观察多因子联合诱导和分步诱导方案诱导大鼠脂肪组织源性干细胞体外分化为神经元样细胞的可行性。
方法：取SD雄性大鼠腹股沟和附睾脂肪垫脂肪组织，酶消化法分离、差速贴壁法纯化脂肪组织源性干细胞。免疫荧光细胞化学法鉴定脂肪组织源性干细胞(CD44，CD49d，CD106)；“鸡尾酒”式神经元诱导培养基对脂肪组织源性干细胞进行诱导分化，并用免疫荧光细胞化学法鉴定(Nestin和NeuN)分化结果。
结果与结论：大鼠腹股沟和附睾脂肪垫脂肪组织内分离、原代培养的脂肪组织源性干细胞形态均一，呈长梭形，细胞核椭圆形，核仁明显，细胞质染色浅，呈旋涡状克隆样生长；CD44和CD49d阳性表达而CD106阴性；诱导培养基内培养6 h后脂肪组织源性干细胞呈锥体形或不规则，有短而细的指状突起；24 h后多数细胞显示类神经元的形态学特征，且Nestin和NeuN阳性。提示肪组织源性干细胞在多因子“鸡尾酒”式培养基内可向神经元方向分化。

关键词: 脂肪组织源性干细胞, 分化, 神经元, 免疫细胞化学, 大鼠

Abstract:

BACKGROUND: Differentiation of adipose tissue-derived stem cells (ADSCs) into neurons has been reported. However, little data are available regarding differential adhesion-purified ADSCs differentiation into neuron-like cells induced by multiple factors.
OBJECTIVE: To explore the feasibility of inducing rat ADSCs into neuron-like cells with a new scheme and medium containing multiple factors.
METHODS: ADSCs were isolated from the fat pads in groins and epididymis of male Sprague Dawley rats by enzyme digestion, and purified by differential attachment method. Fluorescent immunocytochemistry was used to identify the phenotype of the cells (CD44, CD49d and CD106 expressions). Differentiation of ADSCs into neuron-like cells was induced in cocktail-like neuronal differential medium containing insulin, hydrocortisone, valproic acid and forskolin. Also, fluorescent immunocytochemistry was used to identify the markers of the differentiated cells (Nestin and NeuN).
RESULTS AND CONCLUSION: After sequentially isolated, purified and cultured, the obtained cells presented with long-fusiform shape, with clear nucleoli and light stained cytoplasm. They were positive for CD44 and CD49d, but negative for CD106. The application of NIM resulted in morphological changes of ADSCs during the first 6 hours, shown by pyramidal, irregular or triangular cell body with short, thin finger-like cell processes. After 24 hours of induction, most of the cells developed multiple branched processes resembling those of neurons and positive for Nestin and NeuN. Results showed that ADSCs could be induced to differentiate into neuron-like cells in cocktail-like neuronal differential medium.

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