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Updated: Aug 26, 2025
Analysis of Microglia and Monocyte-derived Macrophages from the Central Nervous System by Flow Cytometry
Published on: June 22, 2017
A robust approach to differentiate human monocyte-derived microglia from peripheral blood mononuclear cells
Hazel Quek1, Carla Cuní-López2, Romal Stewart3
1Mental Health and Neuroscience, QIMR Berghofer Medical Research Institute, Brisbane, QLD 4006, Australia; School of Biomedical Science, The University of Queensland, Brisbane, QLD 4072, Australia; School of Biomedical Sciences, Queensland University of Technology, Brisbane, QLD 4059, Australia.
Abstract:
Microglia are implicated in most neurodegenerative diseases. Here, we present a robust and efficient protocol to differentiate monocyte-derived microglia-like cells (MDMi) from whole blood. The protocol consists of three parts. The first part will describe two methods for PBMC isolation. This will be followed by MDMi differentiation, and lastly, the characterization of MDMi by immunocytochemistry. MDMi can be used to investigate microglial-related responses in various age-related neurodegenerative diseases and can be applied to drug testing on a personalized basis. For complete details on the use and execution of this protocol, please refer to Quek et al. (2022).
Insights
This study introduces a new protocol to create microglia-like cells from blood monocytes. These cells aid in studying neurodegenerative diseases and personalized drug testing.
Area of Science:
- Neuroscience
- Immunology
- Cell Biology
Background:
- Microglia play a crucial role in neurodegenerative diseases.
- Existing methods for studying microglia are limited.
- A need exists for accessible models of microglial function.
Purpose of the Study:
- To present a robust and efficient protocol for differentiating monocyte-derived microglia-like cells (MDMi) from whole blood.
- To enable accessible research into microglial roles in neurodegeneration.
- To facilitate personalized drug testing for neurological conditions.
Main Methods:
- Isolation of peripheral blood mononuclear cells (PBMCs) using two distinct methods.
- Differentiation of PBMCs into monocyte-derived microglia-like cells (MDMi).
- Characterization of differentiated MDMi using immunocytochemistry.
Main Results:
- Successful differentiation of MDMi from whole blood.
- Demonstrated robustness and efficiency of the protocol.
- Established a reliable method for generating microglia-like cells for research.
Conclusions:
- The developed protocol provides a valuable tool for investigating microglial responses in neurodegenerative diseases.
- MDMi offer a promising model for studying disease mechanisms and personalized therapeutics.
- This method can advance research in age-related neurological disorders and drug development.
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