上海交通大学学报（医学版）

›› 2013, Vol. 33 ›› Issue (2): 249-.doi: 10.3969/j.issn.1674-8115.2013.02.025

• 技术与方法 • 上一篇    下一篇

ICR胎鼠原代神经元细胞培养及鉴定

陈雪松1,2, 马 姬1, 薛 燕1, 曾凡一1,2   

  1. 1.上海交通大学医学院 医学科学研究院发育生物学研究室, 上海 200025； 2.上海市儿童医院 卫生部医学胚胎分子生物学重点实验室暨上海市胚胎与生殖工程重点实验室, 上海 200040
  • 出版日期:2013-02-28 发布日期:2013-03-07
  • 通讯作者: 曾凡一, 电子信箱: fzeng@sjtu.edu.cn。
  • 作者简介:陈雪松(1983—), 女, 博士； 电子信箱: xschen1983@gmail.com； 马 姬（1973—）, 女, 实验师, 学士； 电子信箱: maanshan0555@sina.com。
  • 基金资助:

    国家自然科学基金（81125003,30900259）；国家高技术研究发展计划项目（2011AA020116）；上海市科委优秀学术带头人项目（12XD1406500）；上海市科委项目（10140900200）；国家重点基础研究发展计划项目（2010CB945200）

Culture and identification of primary neurons isolated from embryonic ICR mice

CHEN Xue-song1,2, MA Ji1, XUE Yan1, ZENG Fan-yi1,2   

  1. 1.Department of Developmental Biology, Institute of Medical Science, Shanghai Jiaotong University School of Medicine, Shanghai 200025, China； 2.Key Laboratory of Medical Embryo Molecular Biology, the Ministry of Health, and Shanghai Laboratory of Embryo and Reproduction Engineering, Shanghai Children´s Hospital, Shanghai Jiaotong University, Shanghai 200040, China
  • Online:2013-02-28 Published:2013-03-07
  • Supported by:

    National Natural Science Foundation of China, 81125003, 30900259； National High-Tech Research and Development Program of China, 863 Program, 2011AA020116； Shanghai Science and Technology Committee Foundation, 12XD1406500, 10140900200； National Basic Research Program of China, 973 Program, 2010CB945200

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摘要：

目的 建立较理想的胎鼠神经元细胞体外原代培养方法。方法 取13.5 d ICR胎鼠的大脑皮层，经剪碎消化得到单细胞悬液进行培养，观察细胞形态并作PCR及Western blotting等进行神经元相关的基因及蛋白Tuj1和Map2的鉴定。结果 细胞生长状态良好，细胞胞体明显，周围有明亮光晕，细胞突触交错形成网络样，通过PCR及Western blotting验证证明所分离培养的细胞是神经元细胞。结论 本培养方法简单易行，可获得典型和纯度较高的ICR胎鼠大脑皮层神经元细胞。

关键词: 神经元, 原代培养, ICR胎鼠, 疾病模型

Abstract:

Objective To establish a favorable primary culture technique for neurons isolated from embryonic ICR mouse cortical tissues. Methods The cortex of embryonic ICR mice aged 13.5 d was isolated, mechanically dissected and digested, and was proceeded to culture. The morphology of neurons was observed, and PCR and Western blotting were applied to identify the expression of Tuj1 and Map2 gene and protein in neurons. Results Cells grew well, with distinct cell body and surrounding bright halation, and there was typical nerve fiber network of synapses. The isolated and cultured cells were confirmed as neurons by PCR and Western blotting. Conclusion This technique is an easy and practical tool for the primary culture of embryonic ICR mouse cortical neurons with high purity.

Key words: neuron, primary culture, embryonic ICR mouse, disease model