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Updated: May 9, 2026
Culturing Microglia from the Neonatal and Adult Central Nervous System
Published on: August 9, 2013
M-CSF increases proliferation and phagocytosis while modulating receptor and transcription factor expression in adult
Amy M Smith1, Hannah M Gibbons, Robyn L Oldfield
1Department of Pharmacology and Clinical Pharmacology, The University of Auckland, Private Bag 92019, Auckland 1142, New Zealand.
Background:
Microglia are the primary immune cells of the brain whose phenotype largely depends on their surrounding micro-environment. Microglia respond to a multitude of soluble molecules produced by a variety of brain cells. Macrophage colony-stimulating factor (M-CSF) is a cytokine found in the brain whose receptor is expressed by microglia. Previous studies suggest a critical role for M-CSF in brain development and normal functioning as well as in several disease processes involving neuroinflammation.
Methods:
Using biopsy tissue from patients with intractable temporal epilepsy and autopsy tissue, we cultured primary adult human microglia to investigate their response to M-CSF. Mixed glial cultures were treated with 25 ng/ml M-CSF for 96 hours. Proliferation and phagocytosis assays, and high through-put immunocytochemistry, microscopy and image analysis were performed to investigate microglial phenotype and function.
Results:
We found that the phenotype of primary adult human microglia was markedly changed following exposure to M-CSF. A greater number of microglia were present in the M-CSF- treated cultures as the percentage of proliferating (BrdU and Ki67-positive) microglia was greatly increased. A number of changes in protein expression occurred following M-CSF treatment, including increased transcription factors PU.1 and C/EBPβ, increased DAP12 adaptor protein, increased M-CSF receptor (CSF-1R) and IGF-1 receptor, and reduced HLA-DP, DQ, DR antigen presentation protein. Furthermore, a distinct morphological change was observed with elongation of microglial processes. These changes in phenotype were accompanied by a functional increase in phagocytosis of Aβ1-42 peptide.
Conclusions:
We show here that the cytokine M-CSF dramatically influences the phenotype of adult human microglia. These results pave the way for future investigation of M-CSF-related targets for human therapeutic benefit.
Insights
Macrophage colony-stimulating factor (M-CSF) significantly alters adult human microglia, increasing their proliferation and phagocytosis. This cytokine influences microglial phenotype, offering potential therapeutic targets for brain conditions.
Area of Science:
- Neuroscience
- Immunology
- Cell Biology
Background:
- Microglia are the brain's primary immune cells, with phenotypes influenced by their microenvironment.
- Macrophage colony-stimulating factor (M-CSF) is a brain cytokine that binds to receptors on microglia.
- M-CSF plays a role in brain development, function, and neuroinflammatory diseases.
Purpose of the Study:
- To investigate the response of primary adult human microglia to M-CSF.
- To determine how M-CSF affects microglial phenotype and function.
Main Methods:
- Primary adult human microglia were cultured from epilepsy and autopsy tissues.
- Cultures were treated with M-CSF (25 ng/ml) for 96 hours.
- Assays included proliferation, phagocytosis, immunocytochemistry, microscopy, and image analysis.
Main Results:
- M-CSF treatment significantly increased microglial proliferation (BrdU, Ki67 positive).
- Protein expression changed, with increased PU.1, C/EBPβ, DAP12, CSF-1R, and IGF-1R, and decreased HLA-DP/DQ/DR.
- Morphological changes included elongated processes, and phagocytosis of Aβ1-42 peptide increased.
Conclusions:
- The cytokine M-CSF dramatically influences the phenotype of adult human microglia.
- These findings suggest M-CSF-related targets for potential human therapeutic interventions.
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