Christopher J. Bohlen, F. Chris Bennett, Mariko L. Bennett
{"title":"Isolation and Culture of Microglia","authors":"Christopher J. Bohlen, F. Chris Bennett, Mariko L. Bennett","doi":"10.1002/cpim.70","DOIUrl":null,"url":null,"abstract":"<p>Microglia represent 5-10% of cells in the central nervous system and contribute to the development, homeostasis, injury, and repair of neural tissues. As the tissue-resident macrophages of the central nervous system, microglia execute core innate immune functions such as detection of pathogens/damage, cytokine secretion, and phagocytosis. However, additional properties that are specific to microglia and their neural environment are beginning to be appreciated. This article describes approaches for purification of microglia by fluorescence-activated cell sorting using microglia-specific surface markers and for enrichment of microglia by magnetic sorting and immunopanning. Detailed information about culturing primary microglia at various developmental stages is also provided. Throughout, we focus on special considerations for handling microglia and compare the relative strengths or disadvantages of different protocols. © 2018 by John Wiley & Sons, Inc.</p>","PeriodicalId":10733,"journal":{"name":"Current Protocols in Immunology","volume":"125 1","pages":""},"PeriodicalIF":0.0000,"publicationDate":"2018-11-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cpim.70","citationCount":"69","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Current Protocols in Immunology","FirstCategoryId":"1085","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1002/cpim.70","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"Immunology and Microbiology","Score":null,"Total":0}
引用次数: 69
Abstract
Microglia represent 5-10% of cells in the central nervous system and contribute to the development, homeostasis, injury, and repair of neural tissues. As the tissue-resident macrophages of the central nervous system, microglia execute core innate immune functions such as detection of pathogens/damage, cytokine secretion, and phagocytosis. However, additional properties that are specific to microglia and their neural environment are beginning to be appreciated. This article describes approaches for purification of microglia by fluorescence-activated cell sorting using microglia-specific surface markers and for enrichment of microglia by magnetic sorting and immunopanning. Detailed information about culturing primary microglia at various developmental stages is also provided. Throughout, we focus on special considerations for handling microglia and compare the relative strengths or disadvantages of different protocols. © 2018 by John Wiley & Sons, Inc.
小胶质细胞的分离与培养
小胶质细胞占中枢神经系统细胞的5-10%,参与神经组织的发育、稳态、损伤和修复。小胶质细胞作为中枢神经系统的常驻巨噬细胞,具有检测病原体/损伤、细胞因子分泌和吞噬等核心先天免疫功能。然而,小胶质细胞及其神经环境特有的其他特性开始受到重视。本文介绍了利用小胶质细胞特异性表面标记的荧光活化细胞分选纯化小胶质细胞的方法,以及通过磁分选和免疫计划富集小胶质细胞的方法。还提供了在不同发育阶段培养初级小胶质细胞的详细信息。在整个过程中,我们将重点放在处理小胶质细胞的特殊考虑因素上,并比较不同方案的相对优势或劣势。©2018 by John Wiley &儿子,Inc。
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