Cryopreservation of cell-based therapies with enhanced cell viability and therapeutic efficacy.

Abstract

Biobanking has played an important role in advancing cell therapy by enabling long-term preservation of cell function, facilitating product transport, and allowing flexibility in infusion scheduling. However, conventional cryopreservation methods rely on high concentrations of dimethyl sulfoxide (DMSO) and serum, which can lead to toxicity, compromise cell viability, complicate post-thaw handling (such as DMSO removal), require additional quality testing, and ultimately reduce therapeutic efficacy. We present a cryopreservation strategy that uses electroporation to introduce protective sugars intracellularly, termed sugar augmented freezing by electroporation (SAFE). Applied across multiple cell types-including chimeric antigen receptor T cells and stem cells-SAFE improved post-thaw viability and expansion by 1.8-fold and enhanced therapeutic efficacy by two-fold compared with standard cryopreservation. Furthermore, SAFE preserved proliferation-associated proteins and metabolites more effectively. Post-thaw cells maintained, and often exceeded, the functional performance of conventionally preserved cells. This work highlights the importance of optimizing cryogenic parameters to achieve greater post-thaw consistency, scalability, and clinical impact on next-generation cell-based therapies.