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Updated: Jun 3, 2026
Derivation of a Human Brain Organoid with Microglia Development
Published on: January 17, 2025
Protocol to generate microglia-containing cerebral organoids to model HIV neuroinflammation
Janet P Zayas1, David Gagliardi1, Stephanie Gambut1
1Department of Microbial Pathogens and Immunity, Rush University Medical Center, Chicago, IL 60612, USA.
Abstract:
Cerebral organoids (COs) are valuable for studying neurodegenerative diseases and pathogens, but their limited microglia pose challenges. Here, we present a protocol to generate organoids with physiologically relevant microglia (CO-iMs). We describe steps for differentiating induced pluripotent stem cells (iPSCs) into mesoderm, seeding embryoid bodies (EBs), and harvesting hematopoietic progenitor cells (HPCs). We detail procedures for co-culturing HPCs and iPSCs to generate CO-iMs, followed by their infection and treatment. The model provides a platform to study viral infections and neuroinflammation. For complete details on the use and execution of this protocol, please refer to Narasipura et al.1.
Insights
Researchers developed a new method to create cerebral organoids (COs) with abundant microglia. This advanced model aids in studying neuroinflammation and viral infections in the brain.
Area of Science:
- Neuroscience
- Stem Cell Biology
- Immunology
Background:
- Cerebral organoids (COs) are crucial for studying brain diseases.
- Limited microglia in current CO models hinder research on neuroinflammation and infections.
- A need exists for improved models with physiologically relevant microglia.
Purpose of the Study:
- To develop a protocol for generating cerebral organoids with abundant, functional microglia.
- To establish a robust model for investigating neurodegenerative diseases and pathogen interactions.
- To facilitate research on viral infections and neuroinflammation in a human brain model.
Main Methods:
- Induced pluripotent stem cells (iPSCs) were differentiated into mesoderm.
- Embryoid bodies (EBs) were seeded and hematopoietic progenitor cells (HPCs) were harvested.
- HPCs were co-cultured with iPSCs to generate cerebral organoids with microglia (CO-iMs).
Main Results:
- A protocol was established for generating CO-iMs with physiologically relevant microglia.
- The CO-iM model allows for the study of viral infections and neuroinflammation.
- This model offers a platform for advanced research into brain pathologies.
Conclusions:
- The developed protocol successfully generates cerebral organoids with microglia (CO-iMs).
- CO-iMs provide a valuable platform for studying neuroinflammation and viral brain infections.
- This model enhances the study of neurodegenerative diseases and related pathologies.
