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Updated: Jun 10, 2025
Generation of Human Microglia to Combine Them with Retinal Organoids for Improved Disease Modeling
Published on: July 26, 2024
Retinal Organoid Microenvironment Enhanced Bioactivities of Microglia-Like Cells Derived From HiPSCs
Mei-Ling Gao1,2, Tong-Yu Wang1,2, Xin Lin1,2
1The State Key Laboratory of Ophthalmology, Optometry and Visual Science, Wenzhou Medical University, Wenzhou, China.
Purpose:
Microglia-like cells derived from stem cells (iMG) provide a plentiful cell source for studying the functions of microglia in both normal and pathological conditions. Our goal is to establish a simplified and effective method for generating iMG in a precisely defined system. Additionally, we aim to achieve functional maturation of iMG through coculture with retinal organoids.
Methods:
In this study, iMG were produced under precisely defined conditions. They were subjected to LPS and poly IC stimulation. Additionally, we examined distinct phenotypic and functional variances between iMG and HMC3, a commonly used human microglia cell line. To investigate how the retinal cell interaction enhances microglial properties, iMG were cocultured with retinal organoids, producing CC-iMG. We performed RNA sequencing, electrophysiological analysis, and transmission electron microscope (TEM) to examine the maturation of CC-iMG compared to iMG.
Results:
Our results demonstrated that iMG performed immune-responsive profiles closely resembling those of primary human microglia. Compared to HMC3, iMG expressed a higher level of typical microglial markers and exhibited enhanced phagocytic activity. The transcriptomic analysis uncovered notable alterations in the ion channel profile of CC-iMG compared to iMG. Electrophysiological examination demonstrated a heightened intensity of inward- and outward-rectifying K+ currents in CC-iMG. Furthermore, CC-iMG displayed elevated numbers of lysosomes and mitochondria, coupled with increased phagocytic activity.
Conclusions:
These findings contribute to advancing our understanding of human microglial biology, specifically in characterizing and elucidating the functions of CC-iMG, thereby offering an in vitro microglial model for future scientific research and potential clinical applications in cell therapy.
Insights
Induced microglia-like cells (iMG) offer a robust model for studying human microglia. Coculturing iMG with retinal organoids enhances their function and maturation, providing a valuable in vitro tool for research and potential cell therapy applications.
Area of Science:
- Stem cell biology
- Neuroscience
- Immunology
Background:
- Microglia are crucial for central nervous system health and disease.
- Induced microglia-like cells (iMG) offer a scalable alternative to primary microglia.
- Existing human microglia cell lines may not fully recapitulate primary cell functions.
Purpose of the Study:
- To develop a simplified and effective method for generating iMG in a defined system.
- To achieve functional maturation of iMG via coculture with retinal organoids.
- To characterize the phenotypic and functional differences between iMG and a standard cell line.
Main Methods:
- Generation of iMG under precisely defined conditions.
- Stimulation of iMG with LPS and poly IC.
- Coculture of iMG with retinal organoids to create CC-iMG.
- Analysis using RNA sequencing, electrophysiology, and transmission electron microscopy (TEM).
Main Results:
- iMG exhibited immune-responsive profiles similar to primary human microglia.
- iMG showed higher expression of microglial markers and enhanced phagocytosis compared to HMC3 cells.
- CC-iMG displayed altered ion channel profiles, increased K+ currents, and enhanced lysosomal and mitochondrial content, with heightened phagocytic activity.
Conclusions:
- iMG serve as a valuable in vitro model for human microglia research.
- Coculture with retinal organoids (CC-iMG) promotes functional maturation.
- These findings advance understanding of microglial biology and offer potential for cell therapy.
