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Updated: Aug 20, 2025
Isolation of Region-specific Microglia from One Adult Mouse Brain Hemisphere for Deep Single-cell RNA Sequencing
Published on: December 3, 2019
Isolation of Human Microglia from Neuropathologically Diagnosed Cases in the Single-Cell Era
Lih-Fen Lue1, Douglas G Walker2, Suet Theng Beh3
1Human Cell Core for Translational Research, Banner Sun Health Research Institute, Sun City, AZ, USA. emilifen@gmail.com.
Abstract:
This chapter describes the core procedures that we have developed over the last two decades to isolate routinely the microglia from postmortem human brains. The method is suitable for brain slices consisting of both gray and white matter.The ability to concomitantly isolate vascular cells with glial cells provides the opportunity to investigate multiple cell types originating from the same donor. This represents a novel approach for -omics research, with the potential for discovering the shared or distinct molecular features among the glia and vascular cells from the same individual.
Insights
Researchers developed a reliable method to isolate microglia and vascular cells from postmortem human brain tissue. This technique enables comparative molecular studies of glial and vascular cells from the same individual for advanced -omics research.
Area of Science:
- Neuroscience
- Cell Biology
- Molecular Biology
Background:
- Microglia are crucial immune cells in the brain.
- Understanding glial and vascular cell interactions is vital for brain health.
- Previous methods for isolating these cells from human postmortem brains were limited.
Purpose of the Study:
- To describe a robust protocol for routine isolation of microglia from human postmortem brains.
- To enable simultaneous isolation of glial and vascular cells from the same brain sample.
- To facilitate comparative -omics studies of multiple cell types from a single donor.
Main Methods:
- Developed and refined a core procedure over two decades.
- The method is applicable to both gray and white matter brain slices.
- Allows for the concomitant isolation of microglia, other glial cells, and vascular cells.
Main Results:
- Established a reliable technique for isolating microglia from postmortem human brain tissue.
- Successfully isolated multiple cell types, including glial and vascular cells, from the same donor.
- The protocol is suitable for diverse brain regions and tissue types.
Conclusions:
- The described method provides a novel approach for studying cell-specific molecular features.
- Enables comparative -omics research on glial and vascular cells from the same individual.
- Offers new opportunities to investigate shared or distinct molecular characteristics among brain cell types.
