Cargando…

A simplified procedure for isolation of primary murine microglia

There are various approaches in which one can isolate microglia from murine brains, such as immunomagnetic, density gradient, FACS and differential adhesive methods. In this procedure a modified flask-tapping approach was used due to its simplicity and reproducibility. Our protocol requires only a s...

Descripción completa

Detalles Bibliográficos
Autores principales: Scott, Nathan, Witt, Ken, Schober, Joseph M
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Future Science Ltd 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9761776/
https://www.ncbi.nlm.nih.gov/pubmed/36398847
http://dx.doi.org/10.2144/btn-2022-0054
_version_ 1784852746230300672
author Scott, Nathan
Witt, Ken
Schober, Joseph M
author_facet Scott, Nathan
Witt, Ken
Schober, Joseph M
author_sort Scott, Nathan
collection PubMed
description There are various approaches in which one can isolate microglia from murine brains, such as immunomagnetic, density gradient, FACS and differential adhesive methods. In this procedure a modified flask-tapping approach was used due to its simplicity and reproducibility. Our protocol requires only a single step to isolate the microglia from the mixed cell population. Once the microglia were isolated, we characterized cell purity, microglial morphology and phagocytic activity. The single-step protocol, without the need for additional astrocyte or oligodendrocyte separation, allows microglial cells to be used immediately for experimental purposes. The protocol is low-cost and can be performed in any lab with standard cell-culture equipment.
format Online
Article
Text
id pubmed-9761776
institution National Center for Biotechnology Information
language English
publishDate 2022
publisher Future Science Ltd
record_format MEDLINE/PubMed
spelling pubmed-97617762022-12-20 A simplified procedure for isolation of primary murine microglia Scott, Nathan Witt, Ken Schober, Joseph M Biotechniques Reports There are various approaches in which one can isolate microglia from murine brains, such as immunomagnetic, density gradient, FACS and differential adhesive methods. In this procedure a modified flask-tapping approach was used due to its simplicity and reproducibility. Our protocol requires only a single step to isolate the microglia from the mixed cell population. Once the microglia were isolated, we characterized cell purity, microglial morphology and phagocytic activity. The single-step protocol, without the need for additional astrocyte or oligodendrocyte separation, allows microglial cells to be used immediately for experimental purposes. The protocol is low-cost and can be performed in any lab with standard cell-culture equipment. Future Science Ltd 2022-11-18 2022-11 /pmc/articles/PMC9761776/ /pubmed/36398847 http://dx.doi.org/10.2144/btn-2022-0054 Text en © 2022 Nathan Scott, Ken Witt, Joseph M. Schober https://creativecommons.org/licenses/by-nc-nd/4.0/This work is licensed under the Attribution-NonCommercial-NoDerivatives 4.0 Unported License (https://creativecommons.org/licenses/by-nc-nd/4.0/)
spellingShingle Reports
Scott, Nathan
Witt, Ken
Schober, Joseph M
A simplified procedure for isolation of primary murine microglia
title A simplified procedure for isolation of primary murine microglia
title_full A simplified procedure for isolation of primary murine microglia
title_fullStr A simplified procedure for isolation of primary murine microglia
title_full_unstemmed A simplified procedure for isolation of primary murine microglia
title_short A simplified procedure for isolation of primary murine microglia
title_sort simplified procedure for isolation of primary murine microglia
topic Reports
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9761776/
https://www.ncbi.nlm.nih.gov/pubmed/36398847
http://dx.doi.org/10.2144/btn-2022-0054
work_keys_str_mv AT scottnathan asimplifiedprocedureforisolationofprimarymurinemicroglia
AT wittken asimplifiedprocedureforisolationofprimarymurinemicroglia
AT schoberjosephm asimplifiedprocedureforisolationofprimarymurinemicroglia
AT scottnathan simplifiedprocedureforisolationofprimarymurinemicroglia
AT wittken simplifiedprocedureforisolationofprimarymurinemicroglia
AT schoberjosephm simplifiedprocedureforisolationofprimarymurinemicroglia