Complex tissue cryopreservation with novel naturally-inspired cryoprotectants

Cryopreservation of composite tissues and organs is a critical need to alleviate suffering, death and increasing healthcare costs. While bioengineered tissues and organs may help alleviate issues arising due to organ and tissue shortage, they are typically designed with the smallest numbers of cells able to sustain tissue function, thus they cannot support more than minimal cell losses due to cryopreservation. Cryopreservation will be key to making these tissues and organs readily available to patients commercially, as well as improving the supply of existing tissue engineered products on the battlefield as well as vascularized complex tissue and vital organs, important for both military and civil tissue bio-banks. However, the primary problem with current cryopreservation media is low cell survival, severe toxicity with DMSO, biodistribution issues and epigenetic changes to cryopreserved cells due to DMSO-based cryoprotectants, and negative side effects. There are no viable solutions currently on the market with low-to-no toxicity for the cryopreservation of tissues. The objective of this proposal is to develop innovative and commercially viable, bioinspired, CPA cocktails to minimize tissue toxicity as compared to commercial slow freezing and vitrification solutions, and to achieve viable bioengineered tissues and organs after thawing.