RosetteArray^® Platform for Quantitative High-Throughput Screening of Human Neurodevelopmental Risk.

Area of Science:

• Neuroscience
• Developmental Biology
• Stem Cell Biology

Background:

• Neural organoids are valuable tools for studying human neurodevelopmental disorders (NDDs).
• Current limitations include a lack of scalable and quantifiable formats for NDD research and drug discovery.
• Standardized methods are needed to assess complex NDD etiologies.

Purpose of the Study:

• To introduce and validate the RosetteArray platform for standardized neural organoid derivation.
• To demonstrate the platform's utility in screening factors contributing to NDDs and neural tube defects.
• To enable quantitative high-throughput screening (qHTS) for neurodevelopmental risk assessment.

Main Methods:

• Utilized the RosetteArray platform, a 96-well plate assay for micropatterned neural rosette tissue generation.
• Seeded RosetteArrays from cryopreserved human pluripotent stem cells.
• Cultured organoids for 6-8 days and quantified results using AI-based software analysis of immunostained images.

Main Results:

• Successfully generated >9000 singularly polarized neural rosette tissues per plate.
• Demonstrated the platform's effectiveness in screening developmental neurotoxicity.
• Showcased suitability for assessing genetic and environmental risk factors for neural tube defects.

Conclusions:

• The RosetteArray platform provides a standardized, scalable, and quantifiable method for neural organoid research.
• It facilitates high-throughput screening for NDDs, developmental neurotoxicity, and risk factors.
• This platform holds potential for regulatory applications and precision medicine in neurodevelopmental disorders.