The isolation of a highly pure and healthy population of macrophages from the CNS in a short time is essential for studying the role of these cells in injury and disease. Current methods rely either on the use of gradients and/or sorting, processes that result either in impure, reduced viability and/or altered populations of cells. Furthermore, RNA extraction after immunopanning is often difficult. Here, a technique combining both gradient isolation and immunopanning to generate highly pure cultures of primary macrophages isolated from the injured adult CNS in <2 h is described. An optimized protocol of a commercially-available RNA extraction kit is also outlined for the isolation of highly pure RNA. A hybridoma cell-line producing CD11b antibody was used to recover activated (CD11b+) macrophages/microglia from the CNS after injury in <2 h with >95% purity.

中文翻译：

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从成年小鼠脊髓中分离并纯化巨噬细胞/小胶质细胞。

在短时间内从中枢神经系统中分离出高纯度和健康的巨噬细胞群对于研究这些细胞在损伤和疾病中的作用至关重要。当前的方法依赖于使用梯度和/或分类，该过程导致不纯的，降低的生存力和/或改变的细胞群。此外，免疫淘洗后的RNA提取通常很困难。在这里，描述了一种技术，该技术结合了梯度分离和免疫淘选技术，可在2小时内从受伤的成人CNS中分离出高纯度的原代巨噬细胞。还概述了市售RNA提取试剂盒的优化方案，用于分离高纯度RNA。杂交瘤细胞系产生的CD11b抗体被用于从中枢神经系统损伤后的中枢神经系统中恢复活化的（CD11b +）巨噬细胞/小胶质细胞。