Expression of Tmem119/Sall1 and Ccr2/CD69 in FACS-Sorted Microglia- and Monocyte/Macrophage-Enriched Cell Populations After Intracerebral Hemorrhage
Year: 2019
Type: article
Abstract: Activation and polarization of microglia and macrophages are critical events in neuroinflammation and hematoma resolution after intracerebral hemorrhage (ICH). However, distinguishing microglia and monocyte-derived macrophages histologically can be difficult. Although they share most cell surface markers, evidence indicates that the gene regulation and function of these two cell types might be different. Flow cytometry is the gold standard for discriminating between the two cell populations, but it is rarely used in the ICH research field. We developed a flow cytometry protocol to identify and sort microglia and monocyte-derived macrophages from mice that have undergone well-established ICH models induced by collagenase or blood injection. In addition, we combined a recently established magnetic-activated cell separation system that allows eight tissue samples to be assessed together. This protocol can be completed within 5-8 h. Sorted cells are fully preserved and maintain expression of microglia-specific (Tmem119/Sall1) and macrophage-specific (Ccr2/CD69) markers. They retain phagocytic ability, respond to lipopolysaccharide stimulation, and engulf fluorescent latex beads. Thus, this protocol represents a very important tool for researching microglial and monocyte-derived macrophage biologic function after ICH and other brain diseases.
Authors: Qian Li, Xi Lan, Xiaoning Han, Jian Wang
Institutions: Johns Hopkins University, Johns Hopkins Medicine
Language: English
Subfield: Neurology
Field: Neuroscience
Domain: Life Sciences
Open Access status: gold
Estimated APC paid: $2,950
Funders: American Heart Association, Johns Hopkins University, National Institutes of Health, Johns Hopkins Bloomberg School of Public Health, Department of Anesthesiology and Critical Care Medicine, Johns Hopkins Medicine, National Institute of Neurological Disorders and Stroke, National Center for Complementary and Integrative Health
Awards: Neuroprotective effect of flavanol (-) epicatechin after intracerebral hemorrhage, Modulation of microglial polarization by interleukin-10 after intracerebral hemorrhage, Role of Secreted Frizzled-related Protein 3 in Regulation of Inflammation after Intracerebral Hemorrhage, G4998371676, PGE2 EP1 and EP3 receptors as therapeutic targets in intracerebral hemorrhage, Microglia/macrophage polarization after intracerebral hemorrhage, Microglia/macrophage polarization after intracerebral hemorrhage, 20-HETE and hemorrhagic brain injury, Role of Soluble Epoxide Hydrolase in Gray and White Matter Injury after Intracerebral Hemorrhage, G898715038
OpenAlex ID: W2909345676