Using mechanical homogenization to isolate microglia from mouse brain tissue to preserve transcriptomic integrity
STAR Protocols. 2022-12-01; 3(4): 101670
DOI: 10.1016/j.xpro.2022.101670
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Herron S(1), Delpech JC(2), Madore C(2), Ikezu T(3).
Author information:
(1)Department of Pharmacology and Experimental Therapeutics, Boston University School of Medicine, Boston, MA, USA. Electronic address: .
(2)Laboratoire NutriNeuro, UMR 1286, Bordeaux INP, INRAE, University of Bordeaux, Bordeaux, France.
(3)Department of Pharmacology and Experimental Therapeutics, Boston University School of Medicine, Boston, MA, USA; Department of Neuroscience, Mayo Clinic Florida, Jacksonville, FL, USA. Electronic address: .
Numerous approaches have been developed to isolate microglia from the brain, but procedures using enzymatic dissociation at 37°C can introduce drastic transcriptomic changes and confound results from gene expression assays. Here, we present an optimized protocol for microglia isolation using mechanical homogenization. We use Dounce homogenization to homogenize mouse brain tissue into single-cell suspension. We then isolate microglia through Percoll gradient and flow cytometry. Isolated microglia exhibit a gene expression pattern without the changes induced by heated enzymatic digestion. For complete details on the use and execution of this protocol, please refer to Clayton et al. (2021).
Copyright © 2022 The Author(s). Published by Elsevier Inc. All rights reserved.
Conflict of interest statement: Declaration of interests The authors declare no
competing interests.