大鼠脊髓星形胶质细胞培养方法的改良  

A modified method to culture astrocytes from neonatal rat spinal cord

作　　者：詹燕[1] 渠文生[1] 田代实[1] 王伟[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院神经内科,武汉430030

出　　处：《解剖学杂志》2009年第5期660-662,共3页Chinese Journal of Anatomy

基　　金：国家自然科学基金重点项目（30800340,30230140）

摘　　要：目的：改进大鼠脊髓星形胶质细胞的体外培养方法，为研究脊髓星形胶质细胞的生物学作用及脊髓相关疾病提供实验模型。方法：新生1～2d的SD乳鼠，无菌操作下游离整段脊柱，将脊髓从椎管中冲出，机械吹打制成单细胞悬液，差速黏附处理以去除成纤维细胞成分。利用GFAP免疫荧光显色对培养细胞进行鉴定。结果：经原代和传代培养，获得的星形胶质细胞纯化率高达99％以上。结论：采用本实验室改进的方法，操作简便快捷，培养的大鼠脊髓星形胶质细胞生长良好，纯度高，为后续实验对脊髓星形胶质细胞的生物学作用研究奠定实验基础。Abstract Objective： The culture method of astrocytes from spinal cord tissues was modified to provide a more valuable ex- perimental model for research. Under a sterile condition, the spines of 1-2-day-old rats were dissected to subsequently obtain spinal cord by washing with D-Hank＇s. Spinal cord cells were dissociated by blowing and then seeded in culture flask. After purified with differential velocity adherent technique, the astrocytes cultured in the second passage was identified with immunofluorescence staining against glial fibrillary acidic protein （GFAP）. Results： Astrocytes growed in a good condition through culture; after a second subculture, astrocytes were purified to 99%. Conclusion： Modified technique for astrocytes culture described in present study is feasible due to its easy operation and harvest of astrocyte with effective purity, which provide an effective tool for sequential function research of astrocytes in spinal cord diseases.

关 键 词：脊髓 星形胶质细胞 细胞培养 大鼠 

分 类 号：R749.61[医药卫生—神经病学与精神病学] R563.1[医药卫生—临床医学]