Advanced culture and recording system for long-term electrophysiological profiling in human brain organoids

Summary/Abstract Brain organoids, derived from human pluripotent stem cells, represent a groundbreaking model system for basic and applied neuroscience and drug development. A significant unmet need exists for better tools for relating electrophysiological activity in neurons and neural circuits to molecular and cellular events over time. This Phase I project will develop an integrated system for large-scale, chronic extracellular neural recording and stimulation in brain organoids, focusing on innovative organoid assay chambers and 3D neural probes. These devices will enable breakthrough studies to correlate molecular and cellular features with functional neuronal and circuit properties across temporal scales from milliseconds to weeks. This project will be of broad utility across basic neuroscience and translational research. We aim to 1) Develop a biological assay chamber supporting chronic electrophysiology while maintaining organoid health; 2) Develop high-fidelity, 3D neural probes for chronic, extracellular recording and stimulation; and 3) Develop an integrated system for turn-key longitudinal electrophysiological assays in brain organoids in cell biology labs. The proposed electrophysiology system will provide, for the first time, a non-destructive, label-free, and high- resolution longitudinal functional assay for diverse types of brain organoids. The system is designed to be used alongside organoid development protocols, other non-destructive assays, and biological/drug treatments for the broadest applicability and minimal imposition of tradeoffs in standard practice. The integrated hardware and software system will enable longitudinal use in well-maintained organoids and turn- key operation by non-specialists. The primary functional requirement is to achieve stable, reliable, high-fidelity multi-modal neural interfaces (record, electrica) from hundreds of neurons and their neural circuits in a prescribed 3D region of interest of a brain organoid for up to 30 days. The tools derived from this project will permit a deeper understanding of the functional aspects of neural interfacing within brain organoids, driving the development of therapeutic strategies and providing a robust framework for the future of neurodevelopmental and neurodegenerative disease research. This project aligns with NOSI NOT-MH-24-110: SBIR/STTR Program Priorities to Bridge Gaps in Advancing the NIMH Mission.