Analysis of Microglia and Monocyte-derived Macrophages from the Central Nervous System by Flow Cytometry
Analysis of Microglia and Monocyte-derived Macrophages from the Central Nervous System by Flow Cytometry
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DOI：
10.3791/55781
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发表时间：
2017-06-01
影响因子：
1.2
通讯作者：
Delarasse, Cecile
中科院分区：
文献类型：
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作者：
Martin, Elodie;El-Behi, Mohamed;Delarasse, Cecile
许多研究已经证明了免疫细胞，特别是巨噬细胞在中枢神经系统（CNS）病理中的作用。CNS中有两种主要的巨噬细胞群体：（i）小胶质细胞，其是CNS的常驻巨噬细胞并且在胚胎发生期间衍生自卵黄囊祖细胞，以及（ii）单核细胞衍生的巨噬细胞（MDM），其可以在疾病期间浸润CNS并且衍生自骨髓祖细胞。每个巨噬细胞亚群的作用根据所研究的病理学而不同。此外，对于用于这些巨噬细胞亚群的组织学标记物或区分标准没有共识。然而，通过流式细胞术分析CD11b和CD45标志物的表达谱，使我们能够区分小胶质细胞（CD11b（+）CD45（中））和MDM（CD11b（+）CD45（高））。在这个协议中，我们表明，密度梯度离心和流式细胞术分析可以用来表征这些CNS巨噬细胞亚群，并研究这些细胞表达的几个标志物的兴趣，我们最近发表的。因此，这项技术可以进一步了解巨噬细胞在神经系统疾病小鼠模型中的作用，也可以用于评估药物对这些细胞的作用。
Numerous studies have demonstrated the role of immune cells, in particular macrophages, in central nervous system (CNS) pathologies. There are two main macrophage populations in the CNS: (i) the microglia, which are the resident macrophages of the CNS and are derived from yolk sac progenitors during embryogenesis, and (ii) the monocyte-derived macrophages (MDM), which can infiltrate the CNS during disease and are derived from bone marrow progenitors. The roles of each macrophage subpopulation differ depending on the pathology being studied. Furthermore, there is no consensus on the histological markers or the distinguishing criteria used for these macrophage subpopulations. However, the analysis of the expression profiles of the CD11b and CD45 markers by flow cytometry allows us to distinguish the microglia (CD11b(+) CD45(med)) from the MDM (CD11b(+) CD45(high)). In this protocol, we show that the density gradient centrifugation and the flow cytometry analysis can be used to characterize these CNS macrophage subpopulations, and to study several markers of interest expressed by these cells as we recently published. Thus, this technique can further our understanding of the role of macrophages in mouse models of neurological diseases and can also be used to evaluate drug effects on these cells.