中国组织工程研究

中国组织工程研究 ›› 2011, Vol. 15 ›› Issue (36): 6789-6792.doi: 10.3969/j.issn.1673-8225.2011.36.032

• 干细胞基础实验 basic experiments of stem cells • 上一篇    下一篇

大鼠胚胎中脑多巴胺神经元的分离培养与鉴定

魏小兵1，邓兴力1，王应莉2，杨智勇2，李智高2，刘如恩3，李  玉1   

  1. 1昆明医学院第一附属医院神经外科，云南省昆明市 650032
    2云南省第一人民医院干部保健科，云南省昆明市 650032
    3中日友好医院神经外科，北京市 100029
  • 收稿日期:2009-10-15 修回日期:2011-07-30 出版日期:2011-09-03 发布日期:2011-09-03
  • 通讯作者: 李玉，硕士，副教授，副主任医师，硕士研究生导师，昆明医学院第一附属医院神经外科，云南省昆明市650032 liyu650410@xinlang.com
  • 作者简介:魏小兵★，男，1982年生，内蒙古自治区巴彦淖尔市人，汉族，昆明医学院在读硕士，主要从事神经外科方面的研究。 875112608@qq.com 并列第一作者：邓兴力☆，男， 1979年生，云南省昆明市人，汉族，2008年昆明医学院毕业，博士，讲师，现在昆明医学院第一附属医院神经外科并担任教学工作 duetch13071@yahoo.com.cn
  • 基金资助:

    云南省教育厅科学研究基金(09Y0153)，课题名称：大鼠胚胎中脑多巴胺神经元的分离培养与鉴定。

Isolation, culture and identification of rat embryonic mesencephalic dopaminergic neurons

Wei Xiao-bing1, Deng Xing-li1, Wang Ying-li2, Yang Zhi-yong2, Li Zhi-gao2, Liu Ru-en3, Li Yu1   

  1. 1Department of Neurosurgery, the First Affiliated Hospital of Kunming Medical College, Kunming  650032, Yunnan Province, China
    2Department of Cadres Health, First People’s Hospital of Yunnan Province, Kunming  650032, Yunnan Province, China
    3Department of Neurosurgery, China-Japan Friendship Hospital, Beijing  100029, China
  • Received:2009-10-15 Revised:2011-07-30 Online:2011-09-03 Published:2011-09-03
  • Contact: Li Yu, Master, Associate professor, Associate chief physician, Master’s supervisor, Department of Neurosurgery, the First Affiliated Hospital of Kunming Medical College, Kunming 650032, Yunnan Province, China
  • About author:Wei Xiao-bing★, Studying for master’s degree, Department of Neurosurgery, the First Affiliated Hospital of Kunming Medical College, Kunming 650032, Yunnan Province, China 875112608@qq.com Deng Xing-li☆, Doctor, Lecturer, Department of Neurosurgery, the First Affiliated Hospital of Kunming Medical College, Kunming 650032, Yunnan Province, China duetch13071@yahoo.com.cn Wei Xiao-bing and Deng Xing-li contributed equally to this paper.
  • Supported by:

    the Science Research Foundation of Yunnan Education Bureau, No. 09Y0153*

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被引次数

1

摘要：

背景：神经细胞移植治疗是最有希望治愈帕金森病的方法之一。
目的：探讨大鼠胚胎中脑多巴胺神经元分离、培养与鉴定的方法。
方法：解剖分离E14d SD大鼠胚胎中脑组织，制备单细胞悬液，以神经细胞培养液培养，观察其生长情况并进行RT-PCR和免疫组织化学鉴定。
结果与结论：在神经细胞培养液中，细胞生长良好。RT-PCR和免疫组织化学结果显示大多数细胞表达多巴胺神经元特异性分子标志。证实实验成功建立了大鼠胚胎中脑多巴胺神经元分离培养与鉴定的方法。

关键词: 多巴胺, 神经元, 中脑, 大鼠, 细胞培养

Abstract:

BACKGROUND: Nerve cell transplantation therapy is one of the most promising methods to cure Parkinson’s disease.
OBJECTIVE: To investigate the culture, isolation and identification of rat embryo mesencephalic dopaminergic neurons.
METHODS: The mesencephalon was dissected from rat embryo at embryonic day 14 (E14). The brain tissue was triturated into a fine single-cell suspension. The cells were cultured with the medium containing Neurobasal, B27 supplement, β-mercaptoethanol and fetal bovine serum. The cells were identified with RT-PCR as well as immunocytochemistry, respectively.
RESULTS AND CONCLUSION: The cells survived well in the medium. RT-PCR and immunocytochemistry results showed mostly all neurons expressed molecular markers for dopaminergic neurons. The dopaminergic neurons derived from rat embryonic mesencephalon were successfully isolated and cultured, which will provide the foundation for the further research about cell therapy of Parkinson’s disease.

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