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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...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Future Science Ltd
2022
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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 |
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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 |
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