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.


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