目的:改良现有的小胶质细胞纯化分离培养方法,建立稳定、高产的培养模型。方法:选用新生1～2d SD大鼠进行混合胶质细胞培养,然后结合营养剥离法及震摇法纯化分离小胶质细胞;利用CD11b/c(OX42)免疫细胞化学的方法对分离的小胶质细胞纯度进行鉴定。结果:改良的方法可稳定的获得5×104个/培养瓶(25cm2)的小胶质细胞,纯度达到95%,荧光倒置显微镜下观察细胞形态,分离第1 d小胶质细胞呈不规则圆形,折光不均匀,继续培养3-5 d,小胶质细胞逐渐出现单极或多极突起,7 d时部分细胞转为静止状态,呈分枝状。结论:改良的小胶质细胞培养方法产量多,纯度高,为小胶质细胞在中枢神经系统疾病相关研究提供了新的基础。 OBJECTIVE: To improve the existing method of purification and isolation of microglia and to establish a stable and high-yield culture model. Methods: Sprague-Dawley rats were used to culture mixed glial cells for 1 ~ 2 days. Then the microglia cells were isolated and purified by the method of nutrient dissection and shaking. The immunostaining of CD11b / c (OX42) Qualitative purity of cells was identified. Results: The modified method could stably obtain microglial cells of 5 × 104 cells / flask (25cm2) with a purity of 95%. The morphological changes of cells were observed under a fluorescence inverted microscope. The microglial cells isolated on the first day showed irregular circles Shape, refractive non-uniform, continue to train 3-5 d, monopolar or multipolar protrusions gradually emerged microglia, 7 d part of the cells into a quiescent state was branched. Conclusion: The improved microglial cell culture method has high yield and high purity, which provides a new basis for the study of microglial diseases in the central nervous system.