Cell Cryoisolation and Cryopreservation without Any Organic Solvent

Abstract

Abstract Cell isolation and banking face challenges, balancing cryoprotective efficacy, functional preservation, and isolation purity, requiring innovative approaches with specificity, scalability, and native functionality. We developed a transformative platform based on DNA nanohydrogels enabling dual preservation-isolation through aptamer-guided trehalose delivery in a single workflow, achieving organic solvent-free cryopreservation and high-efficiency isolation via freeze–thaw lethality disparity, termed cryoisolation. The nanohydrogels encapsulate trehalose in hydrated 3D networks, while aptamer-functionalized frameworks enable receptor-specific targeting, driving rapid cellular internalization and release within 2 h, yielding post-thaw viability comparable to that of DMSO while reducing genomic instability (67% fewer differentially expressed genes). The platform can be extended to cryopreserve mesenchymal stem cells (MSCs), confirming its applicability to sensitive primary cells without compromising functional integrity. Molecular-recognition-guided delivery creates differential freeze–thaw survival, enabling label-free isolation of target cells with >95% purity. By redesigning aptamer sequences, this modular platform demonstrates universal applicability, transforming conventional freeze–thaw cycles into precision cell-selection processes.