-
Tytuł:
-
Isolation and Phenotyping of Adult Mouse Microglial Cells.
-
Autorzy:
-
Grabert K; The Roslin Institute and R(D)SVS, University of Edinburgh, Edinburgh, UK. .
McColl BW; The Roslin Institute and R(D)SVS, University of Edinburgh, Edinburgh, UK.; Dementia Research Institute, Edinburgh Medical School, University of Edinburgh, Edinburgh, UK.
-
Źródło:
-
Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2018; Vol. 1784, pp. 77-86.
-
Typ publikacji:
-
Journal Article; Research Support, Non-U.S. Gov't
-
Język:
-
English
-
Imprint Name(s):
-
Publication: Totowa, NJ : Humana Press
Original Publication: Clifton, N.J. : Humana Press,
-
MeSH Terms:
-
Brain/*cytology
Cell Culture Techniques/*methods
Cell Separation/*methods
Microglia/*cytology
Animals ; Flow Cytometry ; Mice ; Myelin Sheath/chemistry ; Phagocytosis/drug effects ; Povidone/chemistry ; Silicon Dioxide/chemistry
-
Grant Information:
-
BB/J4243/1 United Kingdom BB_ Biotechnology and Biological Sciences Research Council; MR/L003384/10 United Kingdom MRC_ Medical Research Council; MR/L003384/1 United Kingdom MRC_ Medical Research Council; BB/J004332/1 United Kingdom BB_ Biotechnology and Biological Sciences Research Council; MR/K001744/1 United Kingdom MRC_ Medical Research Council
-
Contributed Indexing:
-
Keywords: CD11b immunomagnetic beads; Enzyme digestion; Microglia; Percoll gradient
-
Substance Nomenclature:
-
65455-52-9 (Percoll)
7631-86-9 (Silicon Dioxide)
FZ989GH94E (Povidone)
-
Entry Date(s):
-
Date Created: 20180516 Date Completed: 20190129 Latest Revision: 20220129
-
Update Code:
-
20240104
-
DOI:
-
10.1007/978-1-4939-7837-3_7
-
PMID:
-
29761389
-
Microglia are the resident macrophages of the central nervous system parenchyma and fulfill crucial roles in brain development, homeostasis, and inflammation. The isolation of a pure microglia population from brain tissue enables the examination of microglial phenotypes without the interference of other cell populations. Microglial extractions from the neonatal brain have been described in various protocols, yet the more established and complex adult mouse brain poses a greater challenge. Here we describe a refined protocol including enzymatic and mechanical dissociation of adult mouse brain tissue and removal of myelin by Percoll density gradient. Microglial cells were subsequently extracted by an immunomagnetic approach. This isolation procedure enables the use of functionally viable cells for various applications such as cell culture, flow cytometry, functional assays including bacteria- or bead-based phagocytosis, stimulation assays, and transcriptome profiling techniques such as qRT-PCR and microarray/RNA sequencing.