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Updated: Aug 7, 2026
Analysis of Microglia and Monocyte-derived Macrophages from the Central Nervous System by Flow Cytometry
Published on: June 22, 2017
Characterization of human monocyte-derived microglia-like cells
Cathie Leone1, Gwenaelle Le Pavec, William Même
1Laboratoire de Neuro-Immuno-Virologie, Service de Neurovirologie UMR E-01 CEA and Université Paris-Sud XI, CRSSA, IFR13 Institut Paris Sud Cytokines, Fontenay-aux Roses, France.
Abstract:
Microglial cells are central to brain immunity and intervene in many human neurological diseases. The aim of this study was to develop a convenient cellular model for human microglial cells, suitable for HIV studies. Microglia derive from the hematogenous myelomonocytic lineage, possibly as a distinct subpopulation but in any case able to invade the CNS, proliferate, and differentiate into ameboid and then ramified microglia in the adult life. We thus attempted to derive microglia-like cells from human monocytes. When cultured with astrocyte-conditioned medium (ACM), monocytes acquired a ramified morphology, typical of microglia. They overexpressed substance P and the calcium binding protein Iba-1 and dimly expressed class II MHC, three characteristics of microglial cells. Moreover, they also expressed a potassium inward rectifier current, another microglia-specific feature. These monocyte-derived microglia-like cells (MDMi) were CD4(+)/CD14(+), evocative of an activated microglia phenotype. When treated with lipopolysaccharide (LPS), MDMi lost their overexpression of substance P, which returned to untreated monocyte-derived macrophage (MDM) level. Compared with MDM, MDMi expressed higher CD4 but lower CCR5 levels; they could be infected by HIV-1(BaL), but produced less virus progeny than MDM did. This model of human microglia may be an interesting alternative to primary microglia for large scale in vitro HIV studies and may help to better understand HIV-associated microgliosis and chronic inflammation in the brain.
Insights
Researchers developed a human microglial cell model from monocytes for HIV research. These monocyte-derived microglia-like cells (MDMi) mimic human microglia and can be infected by HIV-1.
Area of Science:
- Neuroscience
- Immunology
- Cell Biology
Background:
- Microglial cells are crucial for brain immunity and implicated in neurological diseases.
- Human microglial cells are essential for studying brain inflammation and infections like HIV.
- Existing models for human microglia are limited, hindering research, especially for HIV studies.
Purpose of the Study:
- To develop a convenient cellular model of human microglial cells for HIV research.
- To characterize monocyte-derived microglia-like cells (MDMi) and assess their suitability for in vitro HIV studies.
Main Methods:
- Human monocytes were cultured with astrocyte-conditioned medium (ACM) to derive microglia-like cells (MDMi).
- MDMi were characterized by morphology, gene expression (Substance P, Iba-1, MHC class II), and electrophysiology (potassium current).
- MDMi were assessed for CD4/CD14 expression, response to lipopolysaccharide (LPS), and susceptibility to HIV-1(BaL) infection compared to monocyte-derived macrophages (MDM).
Main Results:
- Monocytes cultured with ACM acquired microglial morphology and expressed key microglial markers (Substance P, Iba-1, MHC class II, potassium current).
- MDMi exhibited an activated phenotype (CD4+/CD14+) and responded to LPS by altering Substance P expression.
- MDMi showed higher CD4 and lower CCR5 expression than MDM, were infectable by HIV-1(BaL), but produced less virus progeny.
Conclusions:
- Monocyte-derived microglia-like cells (MDMi) provide a viable in vitro model for human microglia.
- This MDMi model is suitable for large-scale in vitro HIV studies.
- The model aids in understanding HIV-associated microgliosis and chronic brain inflammation.
