A Hybrid 2D/3D Approach for Neural Differentiation Into Telencephalic Organoids and Efficient Modulation of FGF8...

Area of Science:

• Developmental Neuroscience
• Stem Cell Biology
• Organoid Technology

Background:

• Human brain development involves complex cell proliferation, differentiation, and migration, orchestrated by morphogens like FGF8.
• Disruptions in early brain development can lead to neurodevelopmental disorders such as autism and epilepsy.
• Studying human brain development in vitro is challenging due to ethical and technical limitations with fetal tissue.

Purpose of the Study:

• To develop a rapid and efficient hybrid 2D/3D protocol for inducing telencephalic organoids.
• To model key aspects of anterior human brain development, including regional patterning.
• To overcome limitations of existing 3D organoid protocols, such as variability and lack of morphogen pathway manipulation.

Main Methods:

• Utilized human induced pluripotent stem cells (hiPSCs) for 2D neural progenitor (NP) induction using small-molecule inhibitors.
• Dissociated NPs were reaggregated and cultured in spinning mini-bioreactors to form 3D organoids.
• Controlled addition of FGF8 modulated regional identity, promoting distinct brain domain development within organoids.

Main Results:

• Achieved rapid and homogenous production of neural progenitors within 10 days.
• Generated 3D telencephalic organoids with neocortical neurons within one month.
• Demonstrated successful modulation of regional identity and domain formation via FGF8 signaling.

Conclusions:

• The hybrid 2D/3D protocol offers a reproducible and efficient method for studying early human brain development.
• This system facilitates the investigation of cellular and molecular mechanisms underlying telencephalic and non-telencephalic brain development.
• The ability to manipulate signaling pathways enhances the complexity and utility of organoids for disease modeling.