title: Isolation of highly enriched primary human microglia for functional studies.
abstract: Microglia, the resident macrophages of the central nervous system play vital roles in brain homeostasis through clearance of pathogenic material. Microglia are also implicated in neurological disorders through uncontrolled activation and inflammatory responses. To date, the vast majority of microglial studies have been performed using rodent models. Human microglia differ from rodent counterparts in several aspects including their response to pharmacological substances and their inflammatory secretions. Such differences highlight the need for studies on primary adult human brain microglia and methods to isolate them are therefore required. Our procedure generates microglial cultures of >95% purity from both biopsy and autopsy human brain tissue using a very simple media-based culture procedure that takes advantage of the adherent properties of these cells. Microglia obtained in this manner can be utilised for research within a week. Isolated microglia demonstrate phagocytic ability and respond to inflammatory stimuli and their purity makes them suitable for numerous other forms of in vitro studies, including secretome and transcriptome analysis. Furthermore, this protocol allows for the simultaneous isolation of neural precursor cells during the microglial isolation procedure. As human brain tissue is such a precious and valuable resource the simultaneous isolation of multiple cell types is highly beneficial.
authors.affiliation: Department of Pharmacology and Clinical Pharmacology, 1023, Auckland, New Zealand.|Centre for Brain Research, 1023, Auckland, New Zealand.
authors.forename: Justin
authors.initials: J
authors.lastname: Rustenhoven
authors.affiliation: Department of Pharmacology and Clinical Pharmacology, 1023, Auckland, New Zealand.|Centre for Brain Research, 1023, Auckland, New Zealand.|Department of Anatomy with Radiology, 1023, Auckland, New Zealand.
authors.forename: Thomas I-H
authors.initials: TI
authors.lastname: Park
authors.affiliation: Centre for Brain Research, 1023, Auckland, New Zealand.|Auckland City Hospital, 1023, Auckland, New Zealand.
authors.forename: Patrick
authors.initials: P
authors.lastname: Schweder
authors.affiliation: Auckland City Hospital, 1023, Auckland, New Zealand.
authors.forename: John
authors.initials: J
authors.lastname: Scotter
authors.affiliation: Auckland City Hospital, 1023, Auckland, New Zealand.
authors.forename: Jason
authors.initials: J
authors.lastname: Correia
authors.affiliation: Centre for Brain Research, 1023, Auckland, New Zealand.
authors.forename: Amy M
authors.initials: AM
authors.lastname: Smith
authors.affiliation: Centre for Brain Research, 1023, Auckland, New Zealand.
authors.forename: Hannah M
authors.initials: HM
authors.lastname: Gibbons
authors.affiliation: Lab Plus, Auckland City Hospital, Auckland, New Zealand.
authors.forename: Robyn L
authors.initials: RL
authors.lastname: Oldfield
authors.affiliation: Centre for Brain Research, 1023, Auckland, New Zealand.|Auckland City Hospital, 1023, Auckland, New Zealand.
authors.forename: Peter S
authors.initials: PS
authors.lastname: Bergin
authors.affiliation: Centre for Brain Research, 1023, Auckland, New Zealand.|Auckland City Hospital, 1023, Auckland, New Zealand.
authors.forename: Edward W
authors.initials: EW
authors.lastname: Mee
authors.affiliation: Centre for Brain Research, 1023, Auckland, New Zealand.|Department of Anatomy with Radiology, 1023, Auckland, New Zealand.
authors.forename: Richard L M
authors.initials: RL
authors.lastname: Faull
authors.affiliation: Centre for Brain Research, 1023, Auckland, New Zealand.|Department of Anatomy with Radiology, 1023, Auckland, New Zealand.
authors.forename: Maurice A
authors.initials: MA
authors.lastname: Curtis
authors.affiliation: Department of Pharmacology and Clinical Pharmacology, 1023, Auckland, New Zealand.|Centre for Brain Research, 1023, Auckland, New Zealand.
authors.forename: E
authors.initials: E
authors.lastname: Scott Graham
authors.affiliation: Department of Pharmacology and Clinical Pharmacology, 1023, Auckland, New Zealand.|Centre for Brain Research, 1023, Auckland, New Zealand.
authors.forename: Mike
authors.initials: M
authors.lastname: Dragunow
mesh_terms: D015415:Biomarkers; D002469:Cell Separation / Q000379:methods; D002478:Cells, Cultured; D018925:Chemokines / Q000096:biosynthesis; D016207:Cytokines / Q000096:biosynthesis; D005786:Gene Expression Regulation; D006801:Humans; D007150:Immunohistochemistry; D053583:Interleukin-1beta / Q000378:metabolism; D017628:Microglia / Q000166:cytology* / Q000502:physiology*; D016328:NF-kappa B / Q000378:metabolism; D010587:Phagocytosis; D010641:Phenotype; D021381:Protein Transport
chemical_list: D015415:Biomarkers; D018925:Chemokines; D016207:Cytokines; D053583:Interleukin-1beta; D016328:NF-kappa B