Materials strategies and multidisciplinary approaches for low-temperature preservation of 3D cellular systems

Recent advances in tissue engineering have paved the way for sophisticated three-dimensional (3D) cellular systems, including organoids and microphysiological systems, which hold great promise in fields like regenerative medicine, disease modeling, and drug discovery. Despite these advances, the lack of effective preservation strategies for 3D systems continues to hinder their biobanking, large-scale distribution, and practical implementation. This review explores the key principles and challenges involved in the low-temperature preservation of 3D cellular constructs and presents multidisciplinary strategies to overcome them. While cryopreservation techniques have been widely used in single cell levels, yet its translation to 3D multicellular systems is limited by restricted cryoprotectant diffusion, cytotoxicity, and nonuniform thermal transfer within 3D complex architectures. Recent innovations integrating thermodynamics, biophysics, biochemistry, materials science, and bioengineering have focused on better ice formation control, minimizing cryoprotectant agent toxicity, and enhancing heat and mass transfer. Finally, the review discusses emerging materials-based and system-level innovations, including nanowarming, novel cryoprotectants, and microfluidics- or isochoric-based systems, which collectively aim to achieve scalable, reproducible, and clinically relevant preservation of organoids, microphysiological systems, and other tissue-scale systems, bridging the translational gap between cellular cryopreservation and functional tissue preservation.