Protocol for characterization of spatiotemporal network dynamics in cortical and hippocampal assembloids

Area of Science:

• Neuroscience
• Stem Cell Biology
• Developmental Biology

Background:

• Human brain circuitry development is complex and challenging to model.
• Human induced pluripotent stem cells (hiPSCs) offer a promising source for generating brain organoids.

Purpose of the Study:

• To establish a protocol for generating functional neural assembloids from hiPSCs.
• To model human brain circuitry, including excitatory and inhibitory neuronal interactions.
• To enable the study of neuronal network integration and activity.

Main Methods:

• Differentiation of excitatory-predominant hippocampal (Hc) and cortical (Cx) organoids from hiPSCs.
• Fusion of Hc and Cx organoids with inhibitory interneuron-predominant ganglionic eminence (GE) organoids.
• Maintenance of assembloids to promote interneuron migration and network integration.
• Functional assessment using two-photon calcium imaging to measure neuronal activity.

Main Results:

• Successful generation of neural assembloids with integrated excitatory and inhibitory neurons.
• Demonstrated interneuron migration and network integration within the assembloids.
• Quantified neuronal activity and network function using advanced imaging techniques.

Conclusions:

• The developed protocol provides a robust method for creating human neural assembloids.
• These assembloids serve as a valuable model for studying human brain circuitry and development.
• This model system facilitates research into neurological disorders and therapeutic interventions.