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In vitro models of microglia: a comparative study
Zoe Woolf1,2, Taylor J Stevenson1,2, Kevin Lee2,3,4
1Department of Pharmacology, The University of Auckland, Auckland, New Zealand.
Abstract:
Microglia perform key homeostatic functions to protect the central nervous system (CNS). However, in many brain disorders their protective functions are abrogated, contributing to disease progression. Therefore, studies of microglial function are critical to developing treatments for brain disorders. Different in vitro microglia models have been established, including primary human and rodent cells, induced pluripotent stem cell (iPSC)-derived models, and immortalised cell lines. However, a direct comparative analysis of the phenotypic and functional characteristics of these models has not been undertaken. Accurate modelling of human microglia in vitro is critical for ensuring the translatability of results from the bench to the brain. Therefore, our study aimed to characterise and compare commonly utilised in vitro microglia models. We assessed four established microglia models: primary human microglia, human iPSC-derived microglia, the human microglial clone 3 (HMC3) cell line, and primary mouse microglia, with primary human brain pericytes acting as a negative control. Primary human microglia, iPSC-derived microglia, and mouse microglia stained positive for myeloid-cell markers (Iba1, CD45 and PU.1), while HMC3 cells only stained positive for mural-cell markers (PDGFRβ and NG2). Distinct secretomes were observed in all cell models in response to inflammatory treatment, with iPSC-derived microglia showing the most significant inflammatory secretions. Notably, nitric oxide was only secreted by mouse microglia. Although all cell types exhibited phagocytic capacity, primary human microglia and iPSC-derived microglia displayed significantly higher levels of phagocytosis. Overall, comparative analysis revealed notable differences between human microglia, iPSC-derived microglia, HMC3 cells and mouse microglia. Such differences should be considered when using these models to study human brain diseases. Experimental findings obtained from mouse models or cell lines should ultimately be cross validated to ensure the translatability of results to the human condition.
Insights
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:
- 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.
