Isolation of Microglia and Immune Infiltrates from Mouse and Primate Central Nervous System
Isolation of Microglia and Immune Infiltrates from Mouse and Primate Central Nervous System
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DOI：
10.1007/978-1-4939-6786-5_23
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发表时间：
2017-01-01
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影响因子：
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通讯作者：
Eggen, Bart J. L.
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文献类型：
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作者：
Galatro, Thais F.;Vainchtein, Ilia D.;Eggen, Bart J. L.
小胶质细胞是中枢神经系统（CNS）的先天性免疫细胞，在维持组织稳态、提供神经支持和神经保护方面发挥重要作用。小胶质细胞不断地观察它们的环境，并迅速对稳态扰动做出反应。小胶质细胞越来越多地参与神经病理学和神经退行性疾病，如阿尔茨海默病、帕金森病和胶质瘤进展。在这里，我们描述了一个详细的小胶质细胞和免疫浸润的分离方案，优化了大量的死后组织从人类和恒河猴，以及较小的组织量从小鼠大脑和脊髓，产生高度纯化的小胶质细胞群体（高达98%的纯度）。这种急性分离方案是基于机械解离和两步密度梯度纯化，然后进行荧光激活细胞分选（FACS）以获得纯的小胶质细胞和免疫浸润群体。
Microglia are the innate immune cells of the central nervous system (CNS) and play an important role in the maintenance of tissue homeostasis, providing neural support and neuroprotection. Microglia constantly survey their environment and quickly respond to homeostatic perturbations. Microglia are increasingly implicated in neuropathological and neurodegenerative conditions, such as Alzheimer's disease, Parkinson's disease, and glioma progression. Here, we describe a detailed isolation protocol for microglia and immune infiltrates, optimized for large amounts of post mortem tissue from human and rhesus macaque, as well as smaller tissue amounts from mouse brain and spinal cord, that yield a highly purified microglia population (up to 98 % purity). This acute isolation protocol is based on mechanical dissociation and a two-step density gradient purification, followed by fluorescence-activated cell sorting (FACS) to obtain pure microglia and immune infiltrate populations.