(402j) Design of Polymeric Cryoprotectants for Quantitative Post-Thaw Recovery of Mammalian Cells after Storage at –196 °C

Approximately 730,000 annual US deaths are attributed to end-stage organ disease while ca. 50,000 patients are added to the US transplant waiting list annually. The discovery and development of a robust approach to complex tissue cryopreservation could revolutionize regenerative medicine through organ and tissue banking, and lead to consequential advancements that would decrease tissue and organ volatility and increase donor tissue availability. Polymeric cryoprotectants are promising alternatives to traditional small molecule cryoprotectants due to their low cytotoxicity, high glass transition temperature, and low osmolality even at high mass concentrations. We designed and synthesized new polymeric cryoprotectants that achieved nearly quantitative post-thaw survival of suspended mammalian cells. Glass transitions, melting/freezing temperatures, and amount of ice formation were measured using differential scanning calorimetry. The kinetics of cellular water transport during freezing were directly observed on a temperature-controlled microscope. Cytotoxicity and long-term post-thaw cell viability were investigated. Through the design of improved cryoprotectants, we aim to enable cryopreservation of tissue and more complex multicellular networks.