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Updated: Jun 21, 2026
Isolation of Pure Astrocytes and Microglia from the Adult Mouse Spinal Cord For In Vitro Assays and Transcriptomic Studies
Published on: October 20, 2023
Rapid isolation and culture of primary microglia from adult mouse spinal cord
Ping K Yip1, Timothy K Y Kaan, Daniel Fenesan
1Neurorestoration Group, Wolfson CARD, King's College London, Guy's Campus, London Bridge, London SE1 1UL, UK. ping.yip@kcl.ac.uk
Abstract:
Microglia are important in homeostasis and widely considered to have roles in the pathogenesis of conditions such as neuropathic pain and multiple sclerosis. The need to study microglia from the adult spinal cord is essential to further understand the role of these cells in disease pathology. Primary microglia are often prepared from brain tissues obtained from embryonic or perinatal age rodents and the process can take over a week to complete. The protocol in this study provides rapid isolation of microglia from adult spinal cord, allowing immediate availability for experimentation of both ex vivo and in vitro within a few hours. A purity of 99% with little or no neuronal or astrocytic contamination can be achieved. Between 70% and 85% of these adult microglia were in a relatively non-activated state. Functionally, these microglia respond to lipopolysaccharide incubation with increases in both phospho-p38 MAPK and OX42 immunostaining, as well as release of ATP, as compared to un-stimulated microglia. This technique provides a protocol to achieve rapid and efficient extraction of high purity, quiescent and functionally active microglia from adult mouse spinal cord, allowing greater study of adult spinal microglia in physiological and pathophysiological states.
Insights
Researchers developed a rapid method to isolate high-purity microglia from adult mouse spinal cords. This technique enables immediate study of these crucial immune cells in both physiological and disease states.
Area of Science:
- Neuroscience
- Immunology
- Cell Biology
Background:
- Microglia are key immune cells in the central nervous system, crucial for homeostasis.
- Their role in neuropathic pain and multiple sclerosis pathogenesis necessitates studying adult spinal cord microglia.
- Current methods for microglia isolation from adult rodents are time-consuming, often taking over a week.
Purpose of the Study:
- To establish a rapid and efficient protocol for isolating high-purity microglia from adult mouse spinal cords.
- To enable immediate use of isolated microglia for both ex vivo and in vitro experiments.
- To facilitate the study of adult spinal microglia in physiological and pathophysiological conditions.
Main Methods:
- Developed a novel protocol for rapid microglia isolation from adult mouse spinal cords.
- Utilized techniques to ensure high purity (99%) with minimal neuronal or astrocytic contamination.
- Assessed the activation state and functional responses of isolated microglia.
Main Results:
- Achieved rapid isolation of microglia within hours, ready for immediate experimentation.
- Obtained high-purity microglia (99%) with minimal contamination.
- Found 70-85% of isolated microglia were in a quiescent state.
- Demonstrated functional activity: microglia responded to lipopolysaccharide (LPS) with increased phospho-p38 MAPK and OX42 immunostaining, and ATP release.
Conclusions:
- The protocol provides a fast, efficient method for obtaining high-purity, quiescent, and functionally active adult mouse spinal microglia.
- This technique significantly advances the study of microglia in adult spinal cord physiology and disease.
- Enables timely investigation into the role of microglia in neurological disorders.
