Isolation of murine postnatal brain microglia for phenotypic characterization using magnetic cell separation technology

Academic Article

Abstract

  • To shorten the time between brain harvesting and microglia isolation, and characterization, we utilized the MACS® neural dissociation kit followed by OctoMACS® CD11b magnetic bead isolation technique to positively select for brain microglia expressing the pan-microglial marker CD11b, a key subunit of the membrane attack complex (MAC). This protocol yields a viable and highly pure (>95%) microglial population of approximately 500,000 cells per pup that is amenable for in vitro characterization within hours or days after being harvested from brain tissue. Primary microglia from C57Bl/6 mice were plated for next-day analyses of morphology and cellular markers by immunocytochemistry or for analysis of gene expression under resting or LPS-stimulated conditions. The ease of isolation enables investigators to perform molecular and cellular analyses without having to wait 1-2 weeks to isolate microglia by conventional methods involving mechanical agitation to dislodge these from astrocyte beds. © 2013 Springer Science+Business Media New York.
  • Authors

    Published In

    Digital Object Identifier (doi)

    Author List

  • Harms AS; Tansey MG
  • Start Page

  • 33
  • End Page

  • 39
  • Volume

  • 1041