*Purpose: Supercooling preservation techniques store a donor organ below 0°C without freezing. This has great advantages in inhibiting metabolism and preserving the organ in comparison to conventional preservation at 4°C. We developed a novel supercooling technique, using a liquid cooling apparatus and novel preservation and perfusion solutions. The purpose of this study was to evaluate the preservation effect of our supercooling preservation technique in a mouse heart transplantation model.

*Methods: Syngeneic heterotopic heart transplantation was performed in three groups of mice: (1) the non-preservation group, in which the cardiac grafts were transplanted immediately after harvesting; (2) the conventional UW (University of Wisconsin solution) group, in which the cardiac grafts were stored in UW solution at 4°C for different periods of time; and (3) the supercooling group, in which the cardiac grafts were stored in a novel supercooling preservation solution at -8°C for different periods of time. The maximal preservation time was investigated. Twenty-four-hour sample data were collected and analyzed to compare the heart graft protection effect of supercooling to conventional UW preservation at 4°C.

*Results: Our technique yielded a stable -8 °C supercooling preservation state, and cardiac graft revival was successfully achieved after preservation for 144 h, long-term survival was observed after preservation for 96 h. The preservation outcomes, including rebeating time, ATP level, myocardial enzyme release, inflammation, neutrophil infiltration, myocardial ultrastructure, myocardial edema, ischemia-reperfusion injury, oxidative stress and myocardial apoptosis, were also improved in the supercooling group.

*Conclusions: Supercooling heart preservation at -8°C greatly prolonged the preservation time and improved the post-transplant outcomes in comparison to conventional 4°C UW preservation. Supercooling preservation is a promising technique for organ preservation.

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