中国医科大学学报

中国医科大学学报

中国医科大学学报 ›› 2009, Vol. 38 ›› Issue (11): 843–.

• 基础医学 • 上一篇    下一篇

大鼠海马神经细胞原代培养及塑料孔板原位鉴定

贾兰玲; 陈相; 庹航行; 贾心善; 陈颖; 李继光   

  1. 中国医科大学基础医学院病理学教研室; 中国医科大学附属第一医院内分泌科
  • 出版日期:2009-12-20 发布日期:2010-04-14
  • 通讯作者: 贾心善 E-mail:XinShanjia@hotmail.com

Primary Culture of Hippocampal Neuron and In Situ Identification in Plastic Plate

  • Online:2009-12-20 Published:2010-04-14

摘要:

目的原代培养大鼠海马神经细胞,探讨简单、有效的塑料孔板原位鉴定方法。方法采用Hiroaki等的方法并做了改良,在塑料孔板里进行胎鼠海马神经细胞原代培养,7d后分别用传统和改良的甲苯胺蓝(TB)染色法、改良的神经细胞特异性烯醇化酶(NSE)染色法对海马神经细胞进行鉴定。结果无血清细胞培养法神经细胞结构特征明显,能形成明显的神经网络结构。传统的TB染色法染色单一,均为淡蓝色;改良的TB染色法红蓝匹配,对比度好;改良的免疫组化NSE染色法胞质阳性。结论无血清培养法是海马神经细胞培养的理想方法,塑料孔板细胞培养原位染色简单易行,对细胞及分子水平研究有重要意义。

关键词: 塑料孔板, 海马神经细胞, 原代培养, 甲苯胺蓝, 神经细胞特异性烯醇化酶

Abstract:

Objective To obtain the primary culture of hippocampal neuron from
      rat brain,and to explore a simple and effective method for in situ
      identification.Methods The hippocampal neurons of fetal rats were cultured
      in plastic plate according to modified Hiroaki method,and then identified
      by traditional and modified toluidine blue(TB)staining and modifed neuron
      specific enolase(NSE)staining 7 days after primary culture.Results We
      cultutred the cells with serum-free medium.The characteristics of neuron
      structurure were obvious,and the neural network was formed.Compared with
      traditional TB staining in which only light blue was shown,the colors of
      modified TB staining included red and blue which were well-contrasted.With
      modified NSE staining,the plasm was stained positively.Conclusion The
      serum-free culture is an ideal approach for the primary culture of
      hippocampal neurons.In situ staining in plastic plate is easy and feasible
      and is of great significance for researches at the celluar and
      molecular level.

Key words: plastic plate, hippocampal neuron, primary culture, toluidine blue, neuron specific enolase

[1] 彭芳,陆永利,杨红卫. 尾核神经元的原代培养及离子通道特性的研究[J]. 中国医科大学学报, 2012, 41(8): 676-678.
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