中国组织工程研究 ›› 2012, Vol. 16 ›› Issue (14): 2563-2566.doi: 10.3969/j.issn.1673-8225.2012.14.019

• 干细胞培养与分化 stem cell culture and differentiation • 上一篇    下一篇

胶原酶和胰酶对胎鼠端脑神经干细胞培养的影响*☆

梁  军1,宋晓玉2,赵富生1,李月珍1,董建将1   

  1. 1牡丹江医学院组织学与胚胎学教研室,黑龙江省牡丹江市157011;2牡丹江心血管病医院,黑龙江省牡丹江市  157011
  • 收稿日期:2011-10-06 修回日期:2011-11-12 出版日期:2012-04-01 发布日期:2012-04-01
  • 通讯作者: 董建将,硕士,讲师,牡丹江医学院组织学与胚胎学教研室,黑龙江省牡丹江市 157011 dongjj101@163.com
  • 作者简介:梁军☆,男,1972年生,黑龙江省牡丹江市人,鲜族,吉林大学白求恩医学部在读博士,副教授,主要从事神经系统疾病治疗的基础研究。
  • 基金资助:

    黑龙江省普通高等学校青年骨干支持项目(1155G60)。

Effects of trypsogen and collagenase on culture of neural stem cells of fetal rat telencephalon  

Liang Jun1, Song Xiao-yu2, Zhao Fu-sheng1, Li Yue-zhen1, Dong Jian-jiang1   

  1. 1Department of Histology and Embryology, Mudanjiang Medical College, Mudanjiang  157011, Heilongjiang Province, China; 2Mudanjiang Cardiovascular Hospital, Mudanjiang  157011, Heilongjiang Province, China
  • Received:2011-10-06 Revised:2011-11-12 Online:2012-04-01 Published:2012-04-01
  • Contact: author: Dong Jian-jiang, Master, Lecturer, Department of Histology and Embryology, Mudanjiang Medical College, Mudanjiang 157011,Heilongjiang Province, China dongjj101@163.com
  • About author:Liang Jun☆, Studying for doctorate, Associate professor, Department of Histology and Embryology, Mudanjiang Medical College, Mudanjiang 157011, Heilongjiang Province, China
  • Supported by:

    the Young Key Scholars Project of Heilongjiang Com-advanced School, No.1155G60*

摘要:

背景:神经干细胞为神经发育和神经功能重建的研究带来了广阔前景,其体外培养已成为神经科学的一个研究基础。
目的:比较胶原酶和胰酶对体外培养神经干细胞的作用。
方法:取孕14 d的SD大鼠,无菌条件下取胎鼠端脑,剪碎后分别用含EDTA的胰酶和Ⅰ型胶原酶消化,无血清培养基培养细胞。
结果与结论:胶原酶消化得到的神经干细胞呈单层贴壁生长,而用胰酶消化得到的则形成聚集体;Nestin免疫荧光鉴定细胞为阳性;获取的神经干细胞纯度大于99%。提示用胶原酶可以简单而快速地获得原代单层化贴壁生长的神经干细胞。
关键词:胰酶;胶原酶;神经干细胞;细胞培养;鉴定;胎鼠
doi:10.3969/j.issn.1673-8225.2012.14.019
 

关键词: 胰酶, 胶原酶, 神经干细胞, 细胞培养, 鉴定, 胎鼠

Abstract:

BACKGROUND: Neural stem cells (NSCs) bring wide prospect for the study of neural development and repairing, and the in vitro culture is foundational to neuroscience.
OBJECTIVE: To contrast the effects of trypsogen and collagenase on culture of NSCs in vitro.
METHODS: The fetal rat telencephalons were isolated from SD rats pregnant for 14 days under sterile conditions, and tissues were divisively treated with trypsogen and typeⅠ collagenase contained EDTA following trituration, and cultured with serum-free medium.
RESULTS AND CONCLUSION: The adherent monoculture NSCs were obtained using collagenase and congeries of cells were obtained using trypsogen. Nestin immunofluorescence identification showed the cells were positive, and the purity was over 99%. The primary adherent monoculture NSCs can be simply and quickly got using collagenase.
 

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