体外少突胶质细胞培养的新方法
投稿时间:2016-08-18  修订日期:2016-09-07  点此下载全文
引用本文:翁超,丁曼,樊尚华,董红娟,卢祖能.体外少突胶质细胞培养的新方法[J].医学研究杂志,2017,46(4):48-51
DOI: 10.11969/j.issn.1673-548X.2017.04.013
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作者单位E-mail
翁超 430060 武汉大学人民医院神经内科  
丁曼 430060 武汉大学人民医院神经内科  
樊尚华 430060 武汉大学人民医院神经内科  
董红娟 430060 武汉大学人民医院神经内科  
卢祖能 430060 武汉大学人民医院神经内科 zunenglu@163.com 
基金项目:湖北省卫生和计划生育委员会重点项目(WJ2015MA007);武汉市科技局应用基础研究计划项目(2015060101010047)
中文摘要:目的 探讨小鼠鼠婴大脑皮质体外少突胶质细胞培养的新方法。方法 取C57/BL6新生小鼠P0~P2(出生后0~2天)的大脑皮质,accummax室温消化10min,然后用含10%FBS的DMEM/F-12高糖培养基体外培养,培养至第7天时,运用巴氏管吹打,从混合培养的细胞中分离纯化少突胶质前体细胞,然后加入促分化培养基促使少突胶质前体细胞分化发育成熟,行细胞免疫荧光进行鉴定。结果 少突胶质前体细胞纯度较高,达到93.3%左右。加入促分化培养基后,少突胶质前体细胞可快速分化发育成熟。结论 该体外少突胶质细胞培养方法较简单,细胞纯度及产量高,对临床脱髓鞘疾病的研究具有很大的应用价值。
中文关键词:少突胶质细胞前体细胞  纯化  增殖  分化
 
New Method for Oligodendrocyte Cell Culture in Vitro
Abstract:Objective To explore a new method for oligodendrocyte cell culture from neonatal mouse cerebral cortex in vitro. Methods We selected P0-P2 (0-2 days after birth) cerebral cortex from C57/BL6 neonatal mice, and digested the tissue at room temperature for 10 minutes through accummax, then used DMEM/F-12 medium with high glucose containing 10% FBS to culture the mixed cells in vitro. After the celles being cultured for 7 days, we used pap tube for pipetting, and purified oligodendrocyte precursor cell (OPCs) from mixed cultured cells. Finally,with oligodendrocyte differentiation medium to promote oligodendrocyte differentiation and maturation, cell lines were identified by immunofluorescence. Results Our approach produced a high yield of purified OPCs, which was about 93.3%.The oligodendrocyte differentiation medium promoted oligodendrocyte differentiation and maturation quickly, when it was added. Conclusion The new method for oligodendrocyte cell culture in vitro is relatively simple with high purity and can produce a high yield of OPCs, which is of great value for clinical demyelinated diseases.
keywords:Oligodendrocyte precursor cells  Purification  Proliferation  Differentiation
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