Advances in Organoid Cryopreservation: Principles, Emerging Strategies, and Engineering Innovations

Organoids, as three-dimensional (3D) self-organizing tissue models, provide physiologically relevant platforms for disease modeling, drug discovery, and regenerative medicine. Effective cryopreservation is essential for their long-term storage, large-scale distribution, and standardized application. This review summarizes the fundamental principles of organoid cryopreservation, including slow-freezing and vitrification strategies, cryoinjury mechanisms, and permeability-based theoretical models, as well as the recent advances in cryoprotective agents. Further progress in organoid preservation is highlighted, focusing on how integrated approaches, such as in situ cryopreservation, hydrogels, and nanotechnology, enhance cryopreservation outcomes. In addition, methods for evaluating the post-thaw structural and functional integrity of organoids are discussed. Finally, we present emerging engineering innovations-including microfluidic control, 3D bioprinting, and biomaterial-assisted systems-that enable precise manipulation and efficient preservation of organoids. Although challenges persist due to the structural complexity and heterogeneity of organoids, the convergence of biophysical principles and engineering technologies offers promising directions toward high-quality and high-throughput organoid preservation.