Human adherent cortical organoids in a multi-well format.

Area of Science:

• Neuroscience
• Stem Cell Biology
• Developmental Biology

Background:

• Human induced pluripotent stem cells (iPSC) are increasingly used to model brain development.
• Existing brain organoid models face challenges in reproducibility and standardization.

Purpose of the Study:

• To develop a novel, reproducible method for generating 3D cortical layers using iPSC-derived neural progenitor cells.
• To characterize the structural and functional maturation of these adherent cortical organoids (ACOs).

Main Methods:

• Seeding frontal cortex-patterned iPSC-derived neural progenitor cells in 384-well plates.
• 8 weeks of differentiation to promote self-organization and 3D cortical layer formation.
• Longitudinal imaging and characterization of organoid development.

Main Results:

• Developed adherent cortical organoids (ACOs) with stereotypical dimensions (3 × 3 × 0.2 mm).
• ACOs contain diverse neural cell types (neurons, astrocytes, oligodendrocytes) and exhibit mature dendritic spines and axonal myelination.
• Demonstrated robust neuronal activity and long-term culture potential (≥10 months).

Conclusions:

• ACOs provide a reproducible, topographically standardized model of human cortical development.
• ACOs overcome limitations of free-floating organoids, such as internal necrosis.
• ACOs are suitable for high-throughput drug discovery, neurotoxicology, and studying brain disorders.