Isolation and Phenotyping of Adult Mouse Microglial Cells

Methods in Molecular Biology
Kathleen Grabert, Barry W McColl

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.

Citations

Aug 8, 2020·Human Vaccines & Immunotherapeutics·Jacob T BeaverIoanna Skountzou
Aug 6, 2020·Frontiers in Cellular Neuroscience·Jinqiang ZhangTao Zhou
Feb 2, 2021·ASN Neuro·Alexander D WalshMichele D Binder
Mar 13, 2021·Frontiers in Immunology·Raffaella MoriniMichela Matteoli
Nov 24, 2021·The Journal of Immunology : Official Journal of the American Association of Immunologists·Patrick O'ConnellAndrea Amalfitano

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