title: Isolation of Microglia and Immune Infiltrates from Mouse and Primate Central Nervous System.
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.
authors.affiliation: Department of Neurology, School of Medicine, University of São Paulo, São Paulo, 01246903, Brazil.
authors.forename: Thais F
authors.initials: TF
authors.lastname: Galatro
authors.affiliation: Department of Neuroscience, Section Medical Physiology, University of Groningen, University Medical Center Groningen, Groningen, 9713 AV, The Netherlands.
authors.forename: Ilia D
authors.initials: ID
authors.lastname: Vainchtein
authors.affiliation: Department of Neuroscience, Section Medical Physiology, University of Groningen, University Medical Center Groningen, Groningen, 9713 AV, The Netherlands.
authors.forename: Nieske
authors.initials: N
authors.lastname: Brouwer
authors.affiliation: Department of Neuroscience, Section Medical Physiology, University of Groningen, University Medical Center Groningen, Groningen, 9713 AV, The Netherlands.
authors.forename: Erik W G M
authors.initials: EWGM
authors.lastname: Boddeke
authors.affiliation: Department of Neuroscience, Section Medical Physiology, University of Groningen, University Medical Center Groningen, Groningen, 9713 AV, The Netherlands. b.j.l.eggen@umcg.nl.
authors.forename: Bart J L
authors.initials: BJL
authors.lastname: Eggen
mesh_terms: D000818:Animals; D000906:Antibodies / Q000737:chemistry; D015703:Antigens, CD / Q000235:genetics / Q000276:immunology; D015415:Biomarkers / Q000378:metabolism; D001921:Brain / Q000276:immunology / Q000473:pathology; D001932:Brain Neoplasms / Q000235:genetics / Q000276:immunology / Q000473:pathology*; D002469:Cell Separation / Q000295:instrumentation / Q000379:methods*; D002499:Centrifugation, Density Gradient / Q000379:methods; D005434:Flow Cytometry / Q000295:instrumentation / Q000379:methods*; D015870:Gene Expression; D005910:Glioma / Q000235:genetics / Q000276:immunology / Q000473:pathology*; D006801:Humans; D008253:Macaca mulatta; D051379:Mice; D017628:Microglia / Q000276:immunology / Q000473:pathology*; D019636:Neurodegenerative Diseases / Q000235:genetics / Q000276:immunology / Q000473:pathology*; D013116:Spinal Cord / Q000276:immunology / Q000473:pathology
chemical_list: D000906:Antibodies; D015703:Antigens, CD; D015415:Biomarkers
keywords: Acute isolation; CNS; Ex vivo; FACS; Glioma; Microglia; Multiple sclerosis; Neuroinflammation