Targeting Oxidative Stress during the Post-Thawing Recovery Window Improves Cryopreservation Outcomes in Human Neural...

Neural organoids serve as critical in vitro models for recapitulating the structural and functional complexities of the human nervous system. However, the transition toward high-throughput drug screening and industrial-scale applications is currently hindered by low post-cryopreservation viability. While traditional research focuses on the freeze-thaw cycle itself, the biological mechanisms driving post-thaw failure remain poorly understood. In this study, we identify distinct cellular vulnerabilities during the recovery phase: mature neurons undergo rapid apoptosis, while neural stem cells exhibit a persistent reduction in proliferative capacity, ultimately leading to organoid growth failure. We demonstrate that these damages are closely associated with the induction of reactive oxygen species. Crucially, post-thaw antioxidant treatment significantly mitigates oxidative stress, restoring proliferative potential and substantially increasing overall organoid survival rates. Our findings suggest that shifting the focus from freezing protocols to targeted post-thaw pharmacological recovery represents a superior strategy for ensuring a reliable, scalable supply of neural organoids for biomedical research.