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Updated: Jul 1, 2026
Magnetic Isolation of Microglial Cells from Neonate Mouse for Primary Cell Cultures
Published on: July 25, 2022
Protocol for isolating and culturing microglia from the adult mouse brain using a magnetic-activated cell sorting
Joohyun Park1, Bhabotosh Barman1, Hyun Kyu Kim1
1Department of Neurology, School of Medicine, University of California, Davis, Stockton Blvd, Sacramento, CA 95817, USA; Institute of Pediatric Reparative Research (IPRM), Shriners Hospital for Children, Sacramento, CA 95817, USA.
Abstract:
Microglia are the resident macrophages of the central nervous system and are highly sensitive to environmental cues, making primary culture challenging. Here, we present a protocol for isolating microglia from adult mouse brains using magnetic-activated cell sorting (MACS). The protocol includes brain sample collection, enzymatic tissue dissociation, step-by-step MACS-based microglia isolation, and the establishment of primary microglial cultures. We validate isolated cells using flow cytometry and quantitative real-time PCR (real-time qPCR) to assess microglial purity.
Insights
This study details a new protocol for isolating microglia, the immune cells of the brain, from adult mice. The method uses magnetic-activated cell sorting (MACS) for high-purity primary microglial cultures.
Area of Science:
- Neuroscience
- Immunology
- Cell Biology
Background:
- Microglia are central nervous system macrophages.
- Primary microglial cultures are challenging due to sensitivity to environmental cues.
Purpose of the Study:
- To present a reliable protocol for isolating microglia from adult mouse brains.
- To enable the establishment of high-purity primary microglial cultures.
Main Methods:
- Magnetic-activated cell sorting (MACS) for microglia isolation.
- Enzymatic tissue dissociation of adult mouse brains.
- Validation of cell purity using flow cytometry and real-time qPCR.
Main Results:
- Successful isolation of microglia from adult mouse brains.
- Establishment of primary microglial cultures with high purity.
- Demonstrated protocol effectiveness through validation techniques.
Conclusions:
- The presented MACS-based protocol provides an effective method for obtaining pure primary microglial cultures.
- This protocol facilitates further research into microglial function and CNS immunity.
