Isolation of highly enriched primary human microglia for functional studies

Scientific Reports
Justin RustenhovenMike Dragunow

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 precurso...Continue Reading

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Citations

Apr 1, 2017·Clinical Infectious Diseases : an Official Publication of the Infectious Diseases Society of America·Fok-Moon LumLisa F P Ng
Aug 21, 2018·Molecular Neurodegeneration·Justin RustenhovenMike Dragunow
Jul 26, 2018·Journal of the American Society of Nephrology : JASN·Yang YangChanglin Mei
May 13, 2018·Journal of Neuroinflammation·Leon C D SmythMike Dragunow
Sep 13, 2019·International Journal of Molecular Sciences·Eleonora Da PozzoClaudia Martini
Jul 22, 2020·Stem Cells Translational Medicine·Christopher S AhujaMichael G Fehlings
Feb 19, 2017·Acta Neuropathologica Communications·Mark R MizeeInge Huitinga
Jul 31, 2020·BMC Cancer·Vanina D HeuserMaria Gardberg
Jul 3, 2019·International Journal of Molecular Sciences·Eryn L WerryMichael Kassiou
Feb 13, 2016·Journal of Neuroinflammation·Justin RustenhovenMike Dragunow
Aug 22, 2018·Frontiers in Cellular Neuroscience·Raissa TimmermanJeffrey J Bajramovic
Jul 21, 2020·Brain Pathology·Nami NakagomiSeiichi Hirota
Aug 8, 2019·Cells·Lih-Fen LueDouglas G Walker
Nov 2, 2020·The Journal of Physiological Sciences : JPS·Tomonori OkazakiShinji Asano
Nov 21, 2020·Brain Communications·Thomas I-H ParkMichael Dragunow
Feb 28, 2021·Communications Biology·Deidre JanssonMike Dragunow
Apr 3, 2021·Acta Neuropathologica Communications·Marina DigregorioBernard Rogister
Sep 14, 2020·Brain, Behavior, and Immunity·Gemma SullivanJames P Boardman
May 25, 2021·STAR Protocols·Zoe WoolfMichael Dragunow
Jul 3, 2021·Materials·Lidija GradišnikTomaž Velnar
Aug 21, 2021·Biochemical Society Transactions·Lorenzo CeccarelliClaudia Martini
Aug 31, 2021·The Neuroscientist : a Review Journal Bringing Neurobiology, Neurology and Psychiatry·James A WisemanThomas I-H Park

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Methods Mentioned

BETA
biopsy
nuclear translocation
confocal microscopy
fluorescence activated cell sorting
FACS
biopsies
scraping

Software Mentioned

MetaXpress®
array
ImageXpress® Micro XLS
Graphpad Prism
FCAP

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