Isolation and culture of pure adult mouse microglia and astrocytes for in vitro characterization and analyses

Mark T Milner; Grace Mep Lawrence; Caroline L Holley; Liviu-Gabriel Bodea; Jürgen Götz; Sabrina S Burgener; Kate Schroder

doi:10.1016/j.xpro.2022.101295

Isolation and culture of pure adult mouse microglia and astrocytes for in vitro characterization and analyses

STAR Protoc. 2022 Apr 11;3(2):101295. doi: 10.1016/j.xpro.2022.101295. eCollection 2022 Jun 17.

Authors

Mark T Milner¹, Grace Mep Lawrence¹, Caroline L Holley¹, Liviu-Gabriel Bodea², Jürgen Götz², Sabrina S Burgener¹, Kate Schroder¹

Affiliations

¹ Institute for Molecular Bioscience and IMB Centre for Inflammation and Disease Research, The University of Queensland, St. Lucia, QLD 4072, Australia.
² Clem Jones Centre for Ageing and Dementia Research, Queensland Brain Institute, The University of Queensland, St. Lucia, QLD 4072, Australia.

Abstract

Microglia and astrocytes are implicated in aging and age-related diseases. Here, we present a protocol to isolate and culture these glia cells from the murine brain. The protocol consists of two parts: magnetic sorting of adult microglia and mechanical/magnetic sorting of adult microglia and astrocytes. We then describe the characterization of these glial cells by flow cytometry and immunohistochemistry. Microglia isolated from aged mice maintain age-related phenotype during culture. These purified glia cells can be applied in ex vivo studies.

Keywords: Cell Biology; Cell culture; Cell isolation; Flow Cytometry/Mass Cytometry; Immunology; Neuroscience; Single Cell.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Astrocytes*
Brain
Flow Cytometry / methods
Mice
Microglia*
Neuroglia