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Updated: Sep 26, 2025
Author Spotlight: Isolation of Glial Cells from the Adult Mouse Spinal Cord
Published on: October 20, 2023
Isolation and culture of pure adult mouse microglia and astrocytes for in vitro characterization and analyses
Mark T Milner1, Grace Mep Lawrence1, Caroline L Holley1
1Institute for Molecular Bioscience and IMB Centre for Inflammation and Disease Research, The University of Queensland, St. Lucia, QLD 4072, Australia.
Abstract:
Microglia and astrocytes are implicated in aging and age-related diseases. Here, we present a protocol to isolate and culture these glia cells from the murine brain. The protocol consists of two parts: magnetic sorting of adult microglia and mechanical/magnetic sorting of adult microglia and astrocytes. We then describe the characterization of these glial cells by flow cytometry and immunohistochemistry. Microglia isolated from aged mice maintain age-related phenotype during culture. These purified glia cells can be applied in ex vivo studies.
Insights
Researchers developed a new protocol to isolate and culture microglia and astrocytes from mouse brains. These aged glial cells retain their age-related characteristics in culture, enabling further ex vivo research.
Area of Science:
- Neuroscience
- Immunology
- Cell Biology
Background:
- Microglia and astrocytes play crucial roles in brain aging and associated diseases.
- Understanding glial cell function in aging requires reliable isolation and culture methods.
Purpose of the Study:
- To present a detailed protocol for isolating and culturing microglia and astrocytes from the adult murine brain.
- To enable the study of age-related glial cell phenotypes in vitro.
Main Methods:
- The protocol involves two main parts: magnetic-activated cell sorting (MACS) for microglia and a combination of mechanical and magnetic sorting for both microglia and astrocytes.
- Characterization of isolated glial cells is performed using flow cytometry and immunohistochemistry.
Main Results:
- The developed protocol successfully isolates and cultures microglia and astrocytes from murine brains.
- Microglia derived from aged mice maintain their age-related phenotype throughout the culture period.
- The purified glial cells are suitable for ex vivo experimental applications.
Conclusions:
- This protocol provides a reliable method for obtaining pure microglia and astrocyte cultures from aged mice.
- The ability to maintain age-related phenotypes in vitro facilitates research into aging and neurodegenerative diseases.
- These purified glial cells are valuable tools for ex vivo investigations of glial biology in aging.
