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.
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This work was supported by grant 1R01 NS072467 (MGT) from NIH/NINDS.
Puntambekar SS, Doose JM, Carson MJ (2008) Microglia: a CNS-specific tissue macrophage. In: Lane TE, Carson M, Bergmann C, Wyss-Coray T (eds) Central nervous system diseases and inflammation. Springer, New York, pp 1–12CrossRefGoogle Scholar
Tansey MG, Wyss-Coray T (2008) Cytokines in CNS inflammation and disease. In: Lane TEC, Carson M, Bergmann C, Wyss-Coray T (eds) Central nervous system diseases and inflammation. Springer, New York, pp 59–106CrossRefGoogle Scholar
Bjorkqvist M, Wild EJ, Thiele J et al (2008) A novel pathogenic pathway of immune activation detectable before clinical onset in Huntington’s disease. J Exp Med 205(8):1869–1877PubMedCrossRefGoogle Scholar
Wyss-Coray T, Mucke L (2002) Inflammation in neurodegenerative disease—a double-edged sword. Neuron 35(3):419–432PubMedCrossRefGoogle Scholar
Marchetti B, Abbracchio MP (2005) To be or not to be (inflamed) – is that the question in anti-inflammatory drug therapy of neurodegenerative disorders? Trends Pharmacol Sci 26(10):517–525PubMedCrossRefGoogle Scholar
McGeer PL, McGeer EG (2004) Inflammation and neurodegeneration in Parkinson’s disease. Parkinsonism Relat Disord 10(Suppl 1):S3–S7PubMedCrossRefGoogle Scholar
Cardona AE, Huang D, Sasse ME et al (2006) Isolation of murine microglial cells for RNA analysis or flow cytometry. Nat Protoc 1(4):1947–1951PubMedCrossRefGoogle Scholar