大脑皮质星形胶质细胞的培养和鉴定

Culture and Identification of Astrocytes from Rat Cortical Tissue

为建立有效的星形胶质细胞体外培养体系,并为体外研究星形胶质细胞的功能奠定基础,本试验于无菌条件下取新生1 d^3 d的SD大鼠的大脑皮质,经1.25 g/L胰酶消化及机械吹打分离细胞,差速贴壁处理以去除成纤维细胞,用含100 mL/L胎牛血清的DMEM/F12培养液培养。之后借助摇床振荡及传代的方法去除其他细胞,并以GFAP-FITC免疫荧光法对所获细胞进行鉴定。结果表明,分离培养的细胞具备典型的星形胶质细胞形态,并表达星形胶质细胞特异性抗原GFAP,纯化率达95%以上。试验表明上述方法可获得符合体外研究要求的星形胶质细胞。

The cultivation method for rat cerebral astrocytes in vitro was studied and modified to settle back ground for further study.Rats aged 1-3 days were used in the experiment.The neonatal rat cerebral cortex was made into suspension by 0.125% trypsin and mechanical dissociation.Differential attachment was used to remove contaminating fibroblasts.The cells were cultured with DMEM/F12 medium containing 10% fetal bovine serum.Then cells of other types were removed by shaker oscillation and repeated passages.The derivation of the cells was identified by immunohistochemical staining with anti-rabbit GFAP.The results showed that isolated cells coincided with the form of astrocytes,expressed GFAP antigen and the proportion of astrocyte was95%.The results inferred that the method described above was reliable in obtaining the high purity astrocytes from neonatal rat cerebral cortex.