中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (49): 9138-.doi: 10.3969/j.issn.1673-8225.2010.49.004

• 干细胞培养与分化 • 上一篇    下一篇

人骨髓间充质干细胞体外定向诱导分化为神经元样细胞

王莹，赵洪昌，赵文静，叶冬霞，李晶，罗速   

  1. 吉林省肝胆病医院医务科，吉林省长春市  130062
  • 出版日期:2010-12-03 发布日期:2010-12-03
  • 作者简介:王莹★，1975年生，吉林省长春市人，汉族，2009年北华大学毕业，硕士，副主任医师，主要从事干细胞治疗终末期肝病的临床应用研究。 xingyuewj2@yahoo.com.cn
  • 基金资助:

    长春市科技发展计划项目(08SF07)。

Directional differentiation of human bone marrow mesenchymal stem cells into neuron-like cells in vitro

Wang Ying, Zhao Hong-chang, Zhao Wen-jing, Ye Dong-xia, Li Jing, Luo Su   

  1. Medical Department, Jilin Liver and Gallbladder Diseases Hospital, Changchun  130062, Jilin Province, China
  • Online:2010-12-03 Published:2010-12-03
  • About author:Wang Ying★, Master, Associate chief physician, Medical Department, Jilin Liver and Gallbladder Diseases Hospital, Changchun 130062, Jilin Province, China xingyuewj2@yahoo.com.cn
  • Supported by:

    the Science and Technology Development Project of Changchun City, No. 08SF07*

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384

被引次数

2

摘要：

背景：文献报道体外诱导骨髓间充质细胞定向分化为神经元样细胞多应用神经生长因子类多肽制剂，选择纯化学诱导剂尚不多见。
目的：建立人骨髓间充质干细胞分离培养体系，体外定向诱导人骨髓间充质干细胞分化为神经元样细胞。
方法：密度梯度离心、贴壁培养法和消化时间控制相结合分离纯化人骨髓间充质干细胞并鉴定，采用β-巯基乙醇和二甲基亚砜诱导分化为神经元样细胞，观察细胞形态，通过尼氏染色、NSE和NF-200免疫细胞化学染色对已分化的神经元样细胞进行鉴定和分化率分析。
结果与结论：分离得到的骨髓间充质干细胞为成纤维样细胞，可见多个核仁，β-巯基乙醇和二甲基亚砜诱导后，间充质干细胞分化为神经元样细胞，伸出较长轴突样和树突样突起且有分支，诱导后的神经元样细胞胞质中存在着深蓝色颗粒状的尼氏小体，NSE、NF-200免疫荧光细胞化学染色均呈阳性，阳性率分别为(85.6±6.7)%和(73.2±5.6)%。结果证实采用密度梯度离心、贴壁培养法和消化时间控制相结合能够成功分离和培养人骨髓间充质干细胞，人骨髓间充质干细胞能够在诱导剂β-巯基乙醇和二甲基亚砜的诱导下体外诱导分化为神经元样细胞。

关键词: 神经细胞, 骨髓间充质干细胞, 分离培养, 诱导分化, &beta, -巯基乙醇, 二甲基亚砜

Abstract:

BACKGROUND: In vitro induced bone marrow mesenchymal stem cells (BMSCs) differentiated into neuron-like cells mainly by polypeptide preparations such as nerve growth factor. Pure chemical inducer is not commonly found.
OBJECTIVE: To establish a system for isolation and culture of BMSCs and induce it directional differentiation into neuron-like cells in vitro.
METHODS: BMSCs were isolated, purified and identified by density gradient centrifugation, adherent culture and digestion time control. BMSCs were induced into neuron-like cells by β-mercaptoethanol and dimethyl sulfoxide. The morphology change was observed. The differentiation ratio analysis of differentiated neuron-like cells was detected by Nissl staining and immunochemical of neuron specific enolase and neurofilament-200.
RESULTS AND CONCLUSION: Separated BMSCs were fibroblast-like cells and they had several nucleoluses. BMSCs could differentiate into neuron-like cells, with the presence of long axons and dendrite-like processes after being induced by β-mercaptoethanol and dimethyl sulfoxide. Both Nissl’s body, neurofilament -200 and neuron specific enolase of neurons after induction were positive and the positive rates of neuron specific enolase and neurofilament-200 were (85.6 ±6.7)% and (73.2 ± 5.6)%, respectively. Results demonstrated that density gradient centrifugation, adherent culture and digestion time control can successfully isolate and culture human BMSCs. Human BMSCs can differentiate into neuron-like cells induced by β-mercaptoethanol and dimethyl sulfoxide in vitro.

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