Related Experiment Video
Updated: Jan 11, 2026
Derivation of a Human Brain Organoid with Microglia Development
Published on: January 17, 2025
Engineering a Perfusion Bioreactor System for hiPSC-Derived Progenitor Co-Culture Capturing Microglial Features in
Catarina M Gomes1,2, Inês de Sá1,2, Margarida Delgado1,2
1iBET, Instituto de Biologia Experimental e Biológica, Oeiras, Portugal.
Abstract:
Microglia are critical regulators of brain homeostasis and immune responses in the central nervous system (CNS). However, existing human-based models fail to reproduce the early and complex microglia-neural cell interactions. The differentiation of human induced pluripotent stem cells (hiPSCs) into specialized cell types offers promising avenues for understanding human development and disease modeling. Herein, a methodology for the differentiation of hiPSC-derived erythromyeloid progenitors (iEMPs) and their 3D co-culture with hiPSC-derived neurospheres were explored, utilizing the Ambr 250 Modular stirred-tank bioreactor (STB) system. The aim of this study was to build a complex co-culture model between iEMP and neurospheres in a scalable and controlled environment. Our results demonstrate that the STB effectively supports the co-culture process, with iEMP integration into the neurospheres, exhibiting cell density, aggregate morphology, and concentration similar to the neurosphere cultures. The co-culture environment induced the upregulation of transcription factors critical for microglial lineage commitment. iEMP-neurospheres displayed a unique secretory profile, releasing proteins involved in extracellular matrix remodeling and neuronal differentiation, essential for microenvironment remodeling. In conclusion, this study underscores the role of iEMPs in CNS development and presents a robust platform for preclinical research.
Insights
Researchers developed a scalable 3D co-culture model using human induced pluripotent stem cells (hiPSCs) to study microglia-neural cell interactions. This model effectively integrates hiPSC-derived erythromyeloid progenitors (iEMPs) with neurospheres, advancing central nervous system (CNS) research.
Area of Science:
- Neuroscience
- Stem Cell Biology
- Developmental Biology
Background:
- Microglia are crucial for central nervous system (CNS) homeostasis and immune responses.
- Existing human models struggle to replicate early and complex microglia-neural cell interactions.
- Human induced pluripotent stem cells (hiPSCs) offer potential for modeling human development and disease.
Purpose of the Study:
- To develop a scalable 3D co-culture system for studying microglia-neural cell interactions.
- To differentiate hiPSC-derived erythromyeloid progenitors (iEMPs) and co-culture them with hiPSC-derived neurospheres.
- To utilize the Ambr 250 Modular stirred-tank bioreactor (STB) system for a controlled and scalable environment.
Main Methods:
- Differentiation of hiPSCs into iEMPs.
- Formation of hiPSC-derived neurospheres.
- 3D co-culture of iEMPs and neurospheres in an Ambr 250 Modular STB system.
- Analysis of cell integration, morphology, transcription factor expression, and secretory profiles.
Main Results:
- The STB system successfully supported iEMP integration into neurospheres, maintaining similar cell density and morphology.
- The co-culture environment promoted microglial lineage commitment through upregulation of key transcription factors.
- iEMP-neurospheres exhibited a distinct secretory profile, releasing factors vital for extracellular matrix remodeling and neuronal differentiation.
Conclusions:
- hiPSC-derived iEMPs play a significant role in CNS development.
- The developed 3D co-culture platform provides a robust and scalable model for preclinical research.
- This model enhances the study of microglia-neural cell interactions in a controlled human-based system.
