Abstract
The development and use of complex cell-based products in clinical and discovery science continues to grow at an unprecedented pace. To this end, cryopreservation plays a critical role, serving as an enabling process, providing on-demand access to biological material, facilitating large scale production, storage, and distribution of living materials. Despite serving a critical role and substantial improvements over the last several decades, cryopreservation often remains a bottleneck impacting numerous areas including cell therapy, tissue engineering, and tissue banking. Studies have illustrated the impact and benefit of controlling cryopreservation-induced delayed-onset cell death (CIDOCD) through various “front end” strategies, such as specialized media, new cryoprotective agents, and molecular control during cryopreservation. While proving highly successful, a substantial level of cell death and loss of cell function remains associated with cryopreservation. Recently, we focused on developing technologies (RevitalICE™) designed to reduce the impact of CIDOCD through buffering the cell stress response during the post-thaw recovery phase in an effort to improve the recovery of previously cryopreserved samples. In this study, we investigated the impact of modulating apoptotic caspase activation, oxidative stress, unfolded protein response, and free radical damage in the initial 24 h post-thaw on overall cell survival. Human hematopoietic progenitor cells in vitro cryopreserved in both traditional extracellular-type and intracellular-type cryopreservation freeze media were utilized as a model cell system to assess impact on survival. Our findings demonstrated that through the modulation of several of these pathways, improvements in cell recovery were obtained, regardless of the freeze media and dimethyl sulfoxide concentration utilized. Specifically, through the use of oxidative stress inhibitors, an average increase of 20% in overall viability was observed. Furthermore, the results demonstrated that by using the post-thaw recovery reagent on samples cryopreserved in intracellular-type media (Unisol™), improvements in overall cell survival approaching 80% of non-frozen controls were attained. While improvements in overall survival were obtained, an assessment on the impact of specific cell subpopulations and functionality remains to be completed. While work remains, these results represent an important step forward in the development of improved cryopreservation processes for use in discovery science, and commercial and clinical settings.
Highlights
At no time in history has cryopreservation played as critical a role as an enabling technology than today
The findings of improved cell survival presented in this study are an important initial step in understanding the potential impact of modulating stress pathway activation associated with cryopreservation during the recovery interval
This study demonstrated that in order to gain an accurate picture of viability, the utilization of more discriminating assays, such as metabolic activity assessment, and extended recovery intervals (24 h vs. 1 h) is necessary
Summary
At no time in history has cryopreservation played as critical a role as an enabling technology than today. From basic research to pharma discovery science, cell therapy, regenerative medicine, tissue engineering, vaccines, endangered species conservation, and organ transplantation, cryopreservation serves as an enabling technology, allowing for a transient halting of biological time [1,2]. According to clinicaltrials.gov, cryopreserved products are used in over 1000 clinical trials involving cell therapy, CAR-T cells, vaccines, stem cells, and engineered tissues worldwide. Low post-thaw yields (recovery), compromised biologic function, and inability for systems to regenerate resulted in many groups shying away from cryopreservation when possible, relegating cryopreservation primarily to the academic and research areas [1,2]. The result was that the development of new life saving biologic products was stifled, as they could only be stored for a few hours to days
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
