Nanotechnology-driven cryopreservation across cells and 3D constructs

Cryopreservation is essential for cell-based research, regenerative medicine, and emerging therapies; however, conventional strategies remain limited by ice-induced damage, osmotic imbalance, and cryoprotectant (CPA)-associated toxicity. This review discusses the mechanisms of cryoinjury, limitations of conventional CPA-based strategies, and emerging approaches, including combinatorial CPA formulations and ice-modulating agents such as antifreeze proteins and synthetic ice recrystallization inhibitors. Building on these advances, we evaluate nanotechnology-enabled platforms for precise and multifunctional cryoprotection, encompassing nanocarrier-mediated intracellular CPA delivery, nanomaterial-based extracellular ice modulation, and uniform volumetric rewarming. We further discuss the challenges and emerging nanotechnology-enabled strategies for preserving three-dimensional cellular constructs, including spheroids and organoids, where diffusion limitations, spatial heterogeneity, and structural complexity require tailored preservation approaches. Together, these advances establish nanotechnology as an integrated platform for next-generation cryopreservation by enabling coordinated control of CPA delivery, ice behavior, and rewarming in regenerative medicine and cell-based therapeutics.