大鼠星形胶质细胞培养方法的改进  

Modification of primary rat astrocyte cultures

作　　者：李会[1] 徐岩[1] 侯筱宇[1] 

机构地区：[1]江苏省脑病生物信息重点实验室,徐州医学院生物化学与分子生物学研究中心,江苏徐州221002

出　　处：《徐州医学院学报》2014年第1期12-14,共3页Acta Academiae Medicinae Xuzhou

基　　金：国家自然科学基金（81173030）;江苏省高校自然科学研究重大项目（11KJA310005）;江苏省高校优势学科建设工程（PAPD）;江苏省青蓝工程

摘　　要：目的 改进星形胶质细胞的体外纯化培养方法,为研究星形胶质细胞的生物学功能提供实验模型.方法 采用新生 SD大鼠,无菌操作取其大脑皮质,用0.025%的胰酶吹打消化成单个细胞,替代传统的0.25%的胰酶水浴消化15 min,单个细胞悬液接种于预先包被好PDL的24孔板或25 cm2的细胞培养瓶中,体外培养9~14天至细胞完全融合并铺满瓶底后进行传代.第2代细胞应用抗GFAP的抗体进行免疫荧光染色来鉴定星形胶质细胞的纯度.结果 免疫荧光染色结果显示这种培养方法分离培养出的星形胶质细胞纯度达到90%以上.结论 采用本实验改进的方法,培养的大鼠星形胶质细胞纯度高、生长状态良好,符合体外研究要求.Objective To establish a modified primary culture system for rat astrocytes to facilitate the biological in- vestigation of the cells. Methods The cerebral cortex was removed from newborn SD rats under sterile conditions, and then broken into single cells by O. 025% trypsase with gentle agitation instead of traditional digestion with 0.25% tryp- sase under a bath at 37~C for 15 minutes. The resultant single cell suspension was seeded onto poly - L - lysine - coated 24 - well plates or 25 cm2 - flasks. After in vitro cultivated for 9 - 14 days, the cells were passaged at nearly 100% con- fluence. The second generation was stained with anti - GFAP antibody to identify the purity of astroeytes through immuno- fluorescence technique. Results The staining results showed more than 90% of the purity of isolated astrocytes. Con- dusion This modified culture technique can be adopted to produce rat astrocytes with higher purity, setting up good foundation for further biological study in astrocytes.

关 键 词：星形胶质细胞 细胞培养 大脑皮质 大鼠 

分 类 号：Q26[生物学—细胞生物学] R329.24[医药卫生—人体解剖和组织胚胎学]