Engineering a Perfusion Bioreactor System for hiPSC-Derived Progenitor Co-Culture Capturing Microglial Features in...

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.