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Isolation and Phenotyping of Adult Mouse Microglial Cells

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https://link.springer.com/protocol/10.1007%2F978-1-4939-7837-3_7
Original languageEnglish
Title of host publicationMacrophages: Methods and Protocols, Methods in Molecular Biology, vol. 1784,
PublisherSpringer
Pages77-86
Number of pages10
Volume1784
ISBN (Print)978-1-4939-7836-6
DOIs
Publication statusPublished - 15 May 2018

Abstract

Microglia are the resident macrophages of the central nervous system parenchyma and fulfill crucial roles in brain development, homeostasis, and inflammation. The isolation of a pure microglia population from brain tissue enables the examination of microglial phenotypes without the interference of other cell populations. Microglial extractions from the neonatal brain have been described in various protocols, yet the more established and complex adult mouse brain poses a greater challenge. Here we describe a refined protocol including enzymatic and mechanical dissociation of adult mouse brain tissue and removal of myelin by Percoll density gradient. Microglial cells were subsequently extracted by an immunomagnetic approach. This isolation procedure enables the use of functionally viable cells for various applications such as cell culture, flow cytometry, functional assays including bacteria- or bead-based phagocytosis, stimulation assays, and transcriptome profiling techniques such as qRT-PCR and microarray/RNA sequencing.

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