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.
CITATION STYLE
Harms, A. S., & Tansey, M. G. (2013). Isolation of murine postnatal brain microglia for phenotypic characterization using magnetic cell separation technology. Methods in Molecular Biology, 1041, 33–39. https://doi.org/10.1007/978-1-62703-520-0_5
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