Validating the efficacy of novel cryopreservation solutions and techniques for applications in cell therapy

Abstract

This study evaluates the efficacy of various cryopreservation solutions in maintaining post-thaw viability and functionality of cell types commonly used in cell therapy. Additionally, it explores the potential benefits of incorporating ADAM-17 inhibitors for maintaining primary human natural killer cell function after cryopreservation and an alternative freezing technique, vitrification, on the survival and function of primary human T cells. Human pluripotent stem cells (hPSCs), primary T cells, and primary natural killer (NK) cells were cryopreserved using both commercially available and novel cryoprotective solutions. Post-thaw assessments included evaluations of short- and long-term viability, phenotypic stability, and functional capacity, tailored to each cell type. Novel cryoprotectants, including CaseCryo DMSO and CaseCryo NON-DMSO provided by CaseBioscience met or exceeded the performance of commercial solutions across these metrics. hPSCs showed the highest long term survival when cryopreserved in CaseCryo DMSO compared to other solutions with a maintenance of normal physiology as shown by differentiation assays. T cells showed similar rates of survival and surface marker expression when cryopreserved in CaseCryo DMSO and CaseCryo NON-DMSO as compared to the currently available industry standard solution, however they showed slightly improved visible morphology after exposure to both the CaseCryo solutions compared to the control. NK cells similarly showed no significant difference in expansion rates and surface marker expression, however donor to donor variability seemed to effect these experiments to a greater degree than the others. Among the ADAM-17 inhibitors tested, GW280264X was most effective in preserving CD16 expression in cryopreserved NK cells. Furthermore, vitrification of T cells emerged as a promising alternative to conventional passive freezing, offering potential advantages in cost and processing time for cell therapy manufacturing.