Innovative Organoid Assessment Incubator—Comprehensive Monitoring of Pressure and Electrophysiology in Brain Organoids with Applications in Electrotherapy Evaluation

Brain organoids, derived from human induced pluripotent stem cells (hiPSCs), serve as advanced three-dimensional in vitro models mimicking human brain architecture and functional. To employ this aspect of brain organoids for neurodevelopmental studies, regenerative medicine, and personalized disease modeling, accurate evaluation of organoids is essential, which necessitates a tracking of their status in multi-perspective over time. However, comprehensive and chronic monitoring remains challenging due to the limitations of conventional analysis techniques, which are often invasive and disrupt the integrity of organoids. Here, we introduce an organoid assessment incubator (OAI) system, a novel platform designed for long-term monitoring of both the cellular status and neural functionality of brain organoids, while maintaining their intact structures. The OAI system employs mass density distribution as a new parameter to evaluate organoid integrity and viability, utilizing ultra-sensitive pressure sensors combined with machine learning-based 3D reconstruction. By integration of liquid metal-based multi-electrode arrays (MEAs) for minimally invasive recording of neural activity within organoids, the OAI system reliably monitored multi-aspects of brain organoids during their maturation. Furthermore, the effects of temporal interference stimulation (TIS) on brain organoids were assessed by the OAI system, demonstrating its efficacy in promoting cell proliferation, neuroplasticity, and reducing central necrosis. By enabling multi-faceted monitoring and therapeutic interventions of brain organoids, the OAI system represents a significant advancement with broad applications in neurological research and therapeutic development.