Isolation of Microglia from Mouse or Human Tissue
Open accessAbstract
Microglia are the innate immune cells of the central nervous system. Although numerous methods have been developed to isolate microglia from the brain, the method of dissociation and isolation can have a profound effect on the function of these highly dynamic cells. Here, we present an optimized protocol to isolate CD11b+ cells (microglia) from mouse or human brain tissue using magnetic bead columns. Isolated microglia can be used to model diseases with neuroinflammatory components for potential therapeutic discoveries. For complete details on the use and execution of this protocol, please refer to Hanamsagar et al. (2017), Rivera et al. (2019), and Edlow et al. (2019).
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Many widely used protocols for microglial isolation continue to include enzymatic digestion, likely because some experimental conditions require greater yield.
“Some experimental conditions may dictate the need for greater yield, especially for older or less-sensitive approaches (including many older bulk RNA-sequencing (RNA-seq) studies or protein-based analyses), which is likely why many widely used and widely cited protocols for microglial isolation continue to include enzymatic digestion4,63,64.”
From Dissection of artifactual and confounding glial signatures by single-cell sequencing of mouse and human brain (2022) · cites this paper as [64]
The Limitations section (‘Warm versus Cold Tissue Dissociation’; page unavailable) reports that warm enzymatic digestion yielded 2–3 times as many cells as cold mechanical dissociation. The Step-by-Step Method Details also include an enzyme digestion mix. These passages support the yield rationale and provide an example of such a protocol, but they do not establish that many widely used protocols include enzymatic digestion or that greater yield is why they do. (LLM, conf=0.84)
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Cites
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Marsh, S. E., Walker, A. J., Kamath, T., Dissing-Olesen, L., Hammond, T. R., de Soysa, T. Y., Young, A. M. H., Murphy, S., Abdulraouf, A., Nadaf, N., et al. (2022) · Nature Neuroscience · doi:10.1038/s41593-022-01022-8 · Cited by 432 · see citing works
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Islam, R., Choudhary, H. H., Zhang, F., Mehta, H., Yoshida, J., Thomas, A. J., & Hanafy, K. (2025) · Scientific Reports · doi:10.1038/s41598-025-96456-y · Cited by 22 · see citing works
- Authors
- Bordt, E. A., Block, C. L., Petrozziello, T., Sadri-Vakili, G., Smith, C. J., Edlow, A. G., & Bilbo, S. D.
- Affiliations
- Harvard University (US)
- Duke University (US)
- Massachusetts General Hospital (US)
- Corresponding author
- Bordt, Evan A.
- Bilbo, Staci D.
- ORCID
- Venue
- STAR Protocols
- Publisher
- Elsevier BV
- Published
- 2020-06
- Volume
- 1
- Issue
- 1
- Pages
- 100035
- ISSN
- 2666-1667 (ISSN-L)
- PMC
- PMC7416840
- OpenAlex
- W3033887425
- Cited by
- 109
- References
- 13
- Citations by year
- 2026: 16 · 2025: 25 · 2024: 19 · 2023: 16 · 2022: 15 · 2021: 15 · 2020: 3
- FWCI
- 4.74
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- Locations
- STAR Protocols (publishedVersion, cc-by-nc-nd)
- PubMed Central (publishedVersion, cc-by-nc-nd)
- PubMed (publishedVersion)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- Funders
- National Institute of Mental Health
- Eunice Kennedy Shriver National Institute of Child Health and Human Development
- National Institute of Environmental Health Sciences
- Topics
- Life Sciences › Neuroscience › Neurology › Neuroinflammation and Neurodegeneration Mechanisms
- Health Sciences › Medicine › Pathology and Forensic Medicine › Multiple Sclerosis Research Studies
- Life Sciences › Neuroscience › Neurology › Neurological Disease Mechanisms and Treatments
- Keywords
- microglia isolation, CD11b+ cells, magnetic bead separation, neuroinflammation, mouse brain, human brain, cell isolation protocol
- MeSH terms
- Animals, Cell Separation, Humans, Microglia, CD11b Antigen, Mice
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