FACS isolation of endothelial cells and pericytes from mouse brain microregions

Crouch, Elizabeth E. and Doetsch, Fiona. (2018) FACS isolation of endothelial cells and pericytes from mouse brain microregions. Nature protocols, 13 (4). pp. 738-751.

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Official URL: https://edoc.unibas.ch/64092/

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The vasculature is emerging as a key contributor to brain function during neurodevelopment and in mature physiological and pathological states. The brain vasculature itself also exhibits regional heterogeneity, highlighting the need to develop approaches for purifying cells from different microregions. Previous approaches for isolation of endothelial cells and pericytes have predominantly required transgenic mice and large amounts of tissue, and have resulted in impure populations. In addition, the prospective purification of brain pericytes has been complicated by the fact that widely used pericyte markers are also expressed by other cell types in the brain. Here, we describe the detailed procedures for simultaneous isolation of pure populations of endothelial cells and pericytes directly from adult mouse brain microregions using fluorescence-activated cell sorting (FACS) with antibodies against CD31 (endothelial cells) and CD13 (pericytes). This protocol is scalable and takes ∼5 h, including microdissection of the region of interest, enzymatic tissue dissociation, immunostaining, and FACS. This protocol allows the isolation of brain vascular cells from any mouse strain under diverse conditions; these cells can be used for multiple downstream applications, including in vitro and in vivo experiments, and transcriptomic, proteomic, metabolomic, epigenomic, and single-cell analysis.
Faculties and Departments:05 Faculty of Science > Departement Biozentrum > Neurobiology > Stem Cell Biology (Doetsch)
UniBasel Contributors:Doetsch, Fiona
Item Type:Article, refereed
Article Subtype:Research Article
Note:Publication type according to Uni Basel Research Database: Journal article
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Last Modified:28 Apr 2020 09:18
Deposited On:28 Apr 2020 09:18

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