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In vitro models of microglia: a comparative study.

Zoe Woolf^1,2, Taylor J Stevenson^1,2, Kevin Lee^2,3,4

¹Department of Pharmacology, The University of Auckland, Auckland, New Zealand.

Scientific Reports

|May 4, 2025

Summary

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This summary is machine-generated.

Comparing in vitro microglia models is crucial for brain disorder research. Human iPSC-derived microglia and primary human microglia show higher phagocytosis and inflammatory responses than cell lines or mouse models.

Area of Science:

Neuroscience
Immunology
Cell Biology

Background:

Microglia are vital for central nervous system (CNS) homeostasis, but their dysfunction contributes to brain disorders.
In vitro models like primary cells, induced pluripotent stem cell (iPSC)-derived cells, and cell lines are used to study microglia.
A direct comparison of these models' phenotypic and functional characteristics is lacking, hindering translatability.

Purpose of the Study:

To characterize and compare commonly used in vitro microglia models.
To assess the phenotypic and functional differences between primary human microglia, human iPSC-derived microglia, HMC3 cell line, and primary mouse microglia.
To evaluate the suitability of these models for studying human brain diseases.

Main Methods:

Keywords:

In vitro models Comparative analysis Human microglial clone 3 (HMC3)Microglia iPSC-derived microglia

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Assessed four microglia models: primary human microglia, human iPSC-derived microglia, HMC3 cell line, and primary mouse microglia.

Used primary human brain pericytes as a negative control.

Analyzed myeloid-cell marker expression (Iba1, CD45, PU.1), mural-cell marker expression (PDGFRβ, NG2), secretomes after inflammatory treatment, and phagocytic capacity.

Main Results:

Primary human microglia, iPSC-derived microglia, and mouse microglia expressed myeloid markers; HMC3 cells expressed mural markers.
Distinct secretomes were observed, with iPSC-derived microglia exhibiting the most significant inflammatory secretions.
Nitric oxide was exclusively secreted by mouse microglia; primary human and iPSC-derived microglia showed higher phagocytosis.

Conclusions:

Significant phenotypic and functional differences exist between human microglia, iPSC-derived microglia, HMC3 cells, and mouse microglia.
These differences must be considered when selecting in vitro models for human brain disease research.
Cross-validation of findings from mouse models or cell lines with human models is essential for ensuring translatability.