Study on isolation and culture of neural stem cells from rat brain

HE Mei; et al

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Study on isolation and culture of neural stem cells from rat brain

更新时间：2024-01-17

- Study on isolation and culture of neural stem cells from rat brain
- Journal of Chongqing Medical University Issue S1, Pages: 44-47(2008)
- 作者机构：
  
  1. 重庆医科大学附属第一医院神经内科
  2. 山西医科大学第一附属医院神经内科
- 作者简介：
- 基金信息：
- DOI：
  CLC： R329
- Published：2008，
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[1]何梅,王学峰,马珊珊,席志芹.大鼠神经干细胞分离和培养的实验研究[J].重庆医科大学学报,2008(S1):44-47.

HE Mei, et al. Study on isolation and culture of neural stem cells from rat brain. [J]. Journal of Chongqing Medical University (S1):44-47(2008)
[1]何梅,王学峰,马珊珊,席志芹.大鼠神经干细胞分离和培养的实验研究[J].重庆医科大学学报,2008(S1):44-47. DOI：

HE Mei, et al. Study on isolation and culture of neural stem cells from rat brain. [J]. Journal of Chongqing Medical University (S1):44-47(2008) DOI：

摘要

目的:探索体外培养大鼠神经干细胞（Neural stem cells

NSCs）的方法和实验条件

建立一种较为可靠的神经干细胞培养方法。方法:分别选取孕14～16dSD胎鼠和新生1d的SD大鼠进行神经干细胞培养

采用单纯机械吹打和机械吹打加酶消化两种方法进行消化

对培养出的细胞进行神经干细胞、神经元、神经胶质细胞的免疫细胞化学和免疫荧光鉴定。结果:用2种来源的大鼠培养出的原代细胞

均表达Nestin

诱导分化后的细胞表达GFAP和MAP-2。结论:大鼠神经干细胞的原代分离与培养的关键是单细胞悬液的获得

消化液浓度太高或消化时间太长

容易因消化过度致细胞损伤甚至死亡。胎鼠来源者神经球形成早

数量多

增殖活力更强

但原代取材过程易被污染。新生鼠来源者鼠源易获得

组织取材过程操作简单

但神经干细胞成球率低。

Abstract

Objective:To establish the method of culturing neural stem cells(NSCs) from rat brain. Methods:E14～16 SD embryonic rat brain and postnatal one-day(P1) rat brain were respectively chose to culture neural stem cells

and two digestive methods that simple mechanical digestion and mechanical combined with chemical digestion were used to determine the best conditions in primary culture.Immunocytochemistry and immunofluorescence were used to identify NSCs

neurons and neuroglial cells. Results:Both the primary cells from two different kinds of rat brains expressed nestin antigen and the differentiated cells expressed GFAP and MAP-2 antigen.Conclusion:The key point of primary NSCs culture was obtaining the monoplast suspension.Overdigestion led to cell injury even death. NSCs from embryonic rat brain had better proliferation ability and could form cell clones earlier and more than NSCs from newborn rat brain

but they were easier to be contaminated. Obtaining newborn rat brain and isolation NSCs from newborn rat brain were easier

but NSCs from newborn rat brain had worse proliferation ability.

关键词

神经干细胞细胞培养免疫荧光免疫生化

Keywords

neural stem cellscell cultureImmunofluorescenceImmunocyto chemistry

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