Isolation of highly enriched primary human microglia for functional studies
| Autor: | Edward W. Mee, Hannah M. Gibbons, John Scotter, Justin Rustenhoven, Richard L.M. Faull, Jason Correia, Thomas I. H. Park, Patrick Schweder, Robyn L. Oldfield, Mike Dragunow, Peter S. Bergin, E. Scott Graham, Amy M. Smith, Maurice A. Curtis |
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| Jazyk: | angličtina |
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Cell type Central nervous system Interleukin-1beta Cell Separation Biology Article Transcriptome 03 medical and health sciences 0302 clinical medicine Phagocytosis Precursor cell medicine Humans Neuroinflammation Cells Cultured Multidisciplinary Microglia NF-kappa B Human brain Immunohistochemistry In vitro Protein Transport 030104 developmental biology medicine.anatomical_structure Phenotype Gene Expression Regulation Immunology Cytokines Chemokines Neuroscience 030217 neurology & neurosurgery Biomarkers |
| Zdroj: | Scientific Reports |
| ISSN: | 2045-2322 |
| DOI: | 10.1038/srep19371 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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