牛肌源性干细胞的培养和鉴定  

Culture and identification of cattle muscle derived stem cells

作　　者：韩颖[1] 孔庆然[1] 尹智[1] 魏延昌[1] 张鑫淼[1] 汪亦男[1] 刘忠华[1] 

机构地区：[1]东北农业大学生命科学学院,哈尔滨150030

出　　处：《东北农业大学学报》2008年第12期47-50,共4页Journal of Northeast Agricultural University

基　　金：东北农业大学创新团队发展计划项目

摘　　要：探讨牛肌源性干细胞(MDSCs)的体外分离、培养、鉴定方法,旨在为牛体细胞核移植研究提供优质的供核细胞。使用胶原酶消化牛骨骼肌,然后采用差速贴壁分离技术纯化获得MDSCs。在倒置显微镜下观察细胞形态,测定生长曲线,分析MDSCs的增殖能力,采用结蛋白(Desmin)、CD34和CD45免疫细胞化学染色对细胞进行鉴定。结果显示,已成功地从牛骨骼肌中分离培养出具有干细胞特征的细胞,该细胞呈Desmin和CD34阳性、CD45阴性。因此,可通过体外原代培养获得高纯度肌源性干细胞。To investigate the methods to separate, culture and identify cattle muscle derived stem cells （MDSCs） as donor cells used for cattle nuclear transfer. To get purified MDSCs, cattle skeletal muscle were digested to single cells by collagenase, and then were separated through differential adhesion. Observed MDSCs morphology under inverted microscope, analyzed reproductive activity by determining growth curves and identified cells by immunostaining with anti-Desmin, CD34 and CD45 antibodies. It successfully isolated cells that possessed properties of stem cells which showed Desmin and CD34 positive and CD45 negative. High purified MDSCs could be isolated through in vitro primary culture.

关 键 词：肌源性干细胞 差速贴壁分离技术 鉴定 

分 类 号：R394.2[医药卫生—医学遗传学] R744[医药卫生—基础医学]