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Generating and characterizing human telencephalic brain organoids from stem cell-derived single neural rosettes
H M Arif Ullah1, Qiju Huang1, Simone Chiola1
1Department of Neurobiology, University of Utah, Salt Lake City, UT, USA.
Nature Protocols
|June 27, 2025
Summary
We developed a method for generating human telencephalic organoids from single neural rosettes. This approach models human brain development and neurodevelopmental disorders, yielding mature neurons for research.
Area of Science:
- Neuroscience
- Developmental Biology
- Stem Cell Biology
Background:
- Human brain development involves complex neural tissue formation.
- Modeling neurodevelopmental disorders requires accurate cellular and structural representations.
Purpose of the Study:
- To present a novel method for generating human telencephalic organoids from single neural rosettes.
- To enable the study of neural cell specification, organization, and modeling of neurodevelopmental disorders.
Main Methods:
- Generating CRISPR-Cas9-engineered human stem cells with SHANK3 deletion.
- Deriving single neural rosettes and differentiating them into telencephalic organoids.
- Utilizing techniques such as single-cell RNA sequencing, immunohistochemistry, and electrophysiology.
Main Results:
- Single neural rosette-derived organoids mimic in vivo neural tube development.
- Organoids show consistent neural cell composition and predictable organization around a lumen.
- Organoids contain mature neurons capable of action potentials and synaptic integration.
Conclusions:
- This protocol provides a robust model for studying human telencephalon development and neurodevelopmental disorders.
- The organoids facilitate research into neural network disruptions and the development of organoid-based biocomputers.
