Abstract
Microglia are the innate immune cells of the central nervous system (CNS) and play an important role in the maintenance of tissue homeostasis, providing neural support and neuroprotection. Microglia constantly survey their environment and quickly respond to homeostatic perturbations. Microglia are increasingly implicated in neuropathological and neurodegenerative conditions, such as Alzheimer’s disease, Parkinson’s disease, and glioma progression. Here, we describe a detailed isolation protocol for microglia and immune infiltrates, optimized for large amounts of post mortem tissue from human and rhesus macaque, as well as smaller tissue amounts from mouse brain and spinal cord, that yield a highly purified microglia population (up to 98 % purity). This acute isolation protocol is based on mechanical dissociation and a two-step density gradient purification, followed by fluorescence-activated cell sorting (FACS) to obtain pure microglia and immune infiltrate populations.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Hoeffel G, Chen J, Lavin Y et al (2015) C-Myb(+) erythro-myeloid progenitor-derived fetal monocytes give rise to adult tissue-resident macrophages. Immunity 42:665–678
Aloisi F (2001) Immune function of microglia. Glia 36:165–179
Ransohoff RM, Perry VH (2009) Microglial physiology: unique stimuli, specialized responses. Annu Rev Immunol 27:119–145
Prinz M, Tay TL, Wolf Y et al (2014) Microglia: unique and common features with other tissue macrophages. Acta Neuropathol 128:319–331
Gosselin D, Link VM, Romanoski CE et al (2014) Environment drives selection and function of enhancers controlling tissue-specific macrophage identities. Cell 159:1327–1340
Lavin Y, Winter D, Blecher-Gonen R et al (2014) Tissue-resident macrophage enhancer landscapes are shaped by the local microenvironment. Cell 159:1312–1326
McMenamin PG, Wealthall RJ, Deverall M et al (2003) Macrophages and dendritic cells in the rat meninges and choroid plexus: three-dimensional localisation by environmental scanning electron microscopy and confocal microscopy. Cell Tissue Res 313:259–269
Chinnery HR, Ruitenberg MJ, McMenamin PG (2010) Novel characterization of monocyte-derived cell populations in the meninges and choroid plexus and their rates of replenishment in bone marrow chimeric mice. J Neuropathol Exp Neurol 69:896–909
Brendecke SM, Prinz M (2015) Do not judge a cell by its cover-diversity of CNS resident, adjoining and infiltrating myeloid cells in inflammation. Semin Immunopathol 37:591–605
Olah M, Raj D, Brouwer N et al (2012) An optimized protocol for the acute isolation of human microglia from autopsy brain samples. Glia 60:96–111
Holtman IR, Raj DD, Miller JA et al (2015) Induction of a common microglia gene expression signature by aging and neurodegenerative conditions: a co-expression meta-analysis. Acta Neuropathol Commun 3:31
Raj DDA, Moser J, van der Pol SMA et al (2015) Enhanced microglial pro-inflammatory response to lipopolysaccharide correlates with brain infiltration and blood–brain barrier dysregulation in a mouse model of telomere shortening. Aging Cell 14:1003–1013
Raj DDA, Jaarsma D, Holtman IR et al (2014) Priming of microglia in a DNA-repair deficient model of accelerated aging. Neurobiol Aging 35:2147–2160
Vainchtein ID, Vinet J, Brouwer N et al (2014) In acute experimental autoimmune encephalomyelitis, infiltrating macrophages are immune activated, whereas microglia remain immune suppressed. Glia 62:1724–1735
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2017 Springer Science+Business Media LLC
About this protocol
Cite this protocol
Galatro, T.F., Vainchtein, I.D., Brouwer, N., Boddeke, E.W.G.M., Eggen, B.J.L. (2017). Isolation of Microglia and Immune Infiltrates from Mouse and Primate Central Nervous System. In: Clausen, B., Laman, J. (eds) Inflammation. Methods in Molecular Biology, vol 1559. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6786-5_23
Download citation
DOI: https://doi.org/10.1007/978-1-4939-6786-5_23
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-6784-1
Online ISBN: 978-1-4939-6786-5
eBook Packages: Springer Protocols