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Microglia pp 17-23 | Cite as

Microglia Isolation from Adult Mouse Brain

  • Jae-Kyung Lee
  • Malú G. Tansey
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1041)

Abstract

Although microglia isolation from embryonic or postnatal mouse brain is possible using a number of different protocols, microglia isolation from adult brain is more challenging and often results in low yields. Here, we describe a protocol to isolate intact microglia from adult mouse brain for functional assays, immunocytochemistry, and/or flow cytometry analysis. This protocol involves enzymatic dissociation in medium supplemented with dispase II, papain, and DNase I followed by mechanical dissociation. Cell separation is achieved via percoll gradients of various densities. Microglia isolated using this protocol is suitable for flow cytometry analysis, RNA isolation for gene expression by real-time PCR or microarrays, and for functional assays including cytokine production, chemotaxis, and phagocytosis.

Key words

Microglia CD11b Inflammatory gene expression Flow cytometry Immunocytochemistry 

Notes

Acknowledgements

This work was supported by grant 1R01 NS072467 (MGT) from NIH/NINDS.

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Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

  • Jae-Kyung Lee
    • 1
  • Malú G. Tansey
    • 1
  1. 1.Emory University School of MedicineAtlantaUSA

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