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...
| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
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Future Science Ltd
2022-12-01
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| Series: | BioTechniques |
| Subjects: | |
| Online Access: | https://www.future-science.com/doi/10.2144/btn-2022-0054 |
| _version_ | 1797980592673914880 |
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| author | Nathan Scott Ken Witt Joseph M Schober |
| author_facet | Nathan Scott Ken Witt Joseph M Schober |
| author_sort | Nathan Scott |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-04-11T05:56:35Z |
| format | Article |
| id | doaj.art-9b4c5bab220041aba694c7b244ddab4b |
| institution | Directory Open Access Journal |
| issn | 0736-6205 1940-9818 |
| language | English |
| last_indexed | 2024-04-11T05:56:35Z |
| publishDate | 2022-12-01 |
| publisher | Future Science Ltd |
| record_format | Article |
| series | BioTechniques |
| spelling | doaj.art-9b4c5bab220041aba694c7b244ddab4b2022-12-22T04:41:52ZengFuture Science LtdBioTechniques0736-62051940-98182022-12-0173627327910.2144/btn-2022-0054A simplified procedure for isolation of primary murine microgliaNathan Scott0Ken Witt1Joseph M Schober21Department of Pharmaceutical Sciences, Southern Illinois University Edwardsville School of Pharmacy, Edwardsville, IL 62026, USA1Department of Pharmaceutical Sciences, Southern Illinois University Edwardsville School of Pharmacy, Edwardsville, IL 62026, USA1Department of Pharmaceutical Sciences, Southern Illinois University Edwardsville School of Pharmacy, Edwardsville, IL 62026, USAThere 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.https://www.future-science.com/doi/10.2144/btn-2022-0054flow cytometryfluorescencemicrogliaprimary celltissue culture |
| spellingShingle | Nathan Scott Ken Witt Joseph M Schober A simplified procedure for isolation of primary murine microglia BioTechniques flow cytometry fluorescence microglia primary cell tissue culture |
| 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 | flow cytometry fluorescence microglia primary cell tissue culture |
| url | https://www.future-science.com/doi/10.2144/btn-2022-0054 |
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