一种改进的星形胶质细胞原代培养方法

摘要

Objective The purpose is to introduce a simplified but effective method for astrocytes culture. Method Newborn Sprague-Dawley (SD) rats (1- day old) were sacrificed by decapitation. The single cell suspension of the cortex of cerebrum was prepared by repeated mechanical pipetting. Differential attachment was used to remove contaminating fibroblasts in the cell suspension. The remaining cells were cultured for 14 days, then were agitated on the rotary shaker (180 rpm, 37 C) to remove microglia. The cells were identified as astrocytes by immunofluorescence staining with anti-GFAP antibody and anti-S100β antibody. Results The rat astrocyte was successfully isolated and the proportion of astrocyte was> 95%. Conclusion This modified cultivation method of astrocyte from rat cerebral tissue is established with high purity, and in a good growth condition.%目的：改进大鼠大脑皮质星形胶质细胞的体外培养，旨在通过较简便的方法获得高纯度形态典型的星形胶质细胞，为研究星形胶质细胞的生物学作用提供实验模型。方法以新生大鼠为星形胶质细胞来源，利用不同规格的吸头相互叠套，逐步机械吹打，使细胞分散，制备单细胞悬液；获得的单细胞悬液用差速黏附处理去除成纤维细胞，原代培养14d后，恒温振荡法去除小胶质细胞等杂质细胞；胶质细胞原纤维酸性蛋白（GFAP）和S100β免疫荧光共染色，鉴定细胞纯度。结果这种改进的星形胶质细胞培养方法，分离培养出星形胶质细胞，阳性率达95%以上。结论采用机械吹打方法建立原代星形胶质细胞培养体系，是一种比较简便、值得推广的可用于体外细胞培养研究工作的有效方法。