Partial freezing of rat livers extends preservation time by 5-fold
Shannon N. Tessier, Reinier J. de Vries, Casie A. Pendexter, Stephanie E. J. Cronin, Sinan Özer, Ehab Hafiz, Siavash Raigani, João Paulo Oliveira–Costa, Benjamin T. Wilks, Manuela Lopera Higuita, Thomas M. van Gulik, O. Berk Usta, Shannon L. Stott, Heidi Yeh, Martin L. Yarmush, Korkut Uygun, Mehmet Toner
Abstract
The limited preservation duration of organs has contributed to the shortage of organs for transplantation. Recently, a tripling of the storage duration was achieved with supercooling, which relies on temperatures between -4 and -6 °C. However, to achieve deeper metabolic stasis, lower temperatures are required. Inspired by freeze-tolerant animals, we entered high-subzero temperatures (-10 to -15 °C) using ice nucleators to control ice and cryoprotective agents (CPAs) to maintain an unfrozen liquid fraction. We present this approach, termed partial freezing, by testing gradual (un)loading and different CPAs, holding temperatures, and storage durations. Results indicate that propylene glycol outperforms glycerol and injury is largely influenced by storage temperatures. Subsequently, we demonstrate that machine perfusion enhancements improve the recovery of livers after freezing. Ultimately, livers that were partially frozen for 5-fold longer showed favorable outcomes as compared to viable controls, although frozen livers had lower cumulative bile and higher liver enzymes.