Deriving early single-rosette brain organoids from human pluripotent stem cells.

Area of Science:

• Neuroscience
• Developmental Biology
• Stem Cell Biology

Background:

• Current brain organoid methods face challenges with structural variability and multiple rosette formations.
• Reproducible and standardized brain organoid models are crucial for studying human brain development and neurological disorders.

Purpose of the Study:

• To develop a novel human brain organoid technique that generates self-organizing, single-rosette cortical organoids (SOSR-COs).
• To establish a reproducible method for creating brain organoids with consistent size and structure at early developmental stages.
• To provide a valuable tool for studying human brain development and modeling neurodevelopmental disorders.

Main Methods:

• Initiated brain organoid formation from a 2D monolayer of human pluripotent stem cells.
• Patterned cells into neuroepithelium using small molecules and differentiated them to dorsal cerebral cortex cells.
• Utilized a 2D to 3D developmental transition approach, mimicking neural plate to neural tube formation.

Main Results:

• Successfully generated self-organizing, single-rosette cortical organoids (SOSR-COs) with reproducible size and structure.
• Observed the development of appropriate progenitor and cortical laminar cell types via immunocytochemistry and single-cell RNA sequencing.
• Demonstrated robust structural phenotypes in SOSR-COs following chemical teratogen exposure and in modeling a genetic neurodevelopmental disorder.

Conclusions:

• The developed SOSR-CO technique offers a standardized and reproducible method for human brain organoid generation.
• This method effectively recapitulates early human brain development and is suitable for disease modeling.
• SOSR-COs provide a powerful platform for investigating neurodevelopmental processes and the impact of genetic or environmental factors.