A Multithermic Machine Perfusion System Capable of Precise Control of Perfusion Parameters and Stepwise Loading of Cryoprotectants for Mutiscale Organ Preservation

Abstract Organ transplantation is the only effective therapeutic modality for end-stage organ diseases. However, short preservation windows of traditional static cold storage (several hours) and donor scarcity limit the development of transplantation. Machine perfusion (MP) has emerged as a superior preservation and repair strategy. It has been demonstrated that a reduction in temperature can further diminish organ metabolic rates, thus minimizing consumptive injury. Currently, machine perfusion devices for organs above 0 °C are available, but those for subzero temperatures and suitable for multiple organs remain underdeveloped. Here, we develop a multithermic machine perfusion (MTMP) system, which enables programmable and precise regulation over a wide temperature range from normothermia (37 °C), hypothermia (4 °C) to supercooling (<0 °C). The engineered platform incorporates a dual-channel temperature control module, a closed-loop perfusion unit with pressure feedback, and a scalable organ chamber adaptable to diverse organs (1–200 cm3). The device demonstrates precise control over temperature, pressure, and flowrate. We successfully perform perfusion experiments on rat hearts, rabbit kidneys and pig kidneys using the designed MTMP system, achieving stable multithermic perfusion preservation. Moreover, the system enabled stepwise cryoprotectant loading, establishing a critical pretreatment platform for organ vitrification. This MTMP system addresses critical gaps in organ preservation technology with its multiscale applicability and compatibility with vitrification pretreatment, providing an engineering foundation for extending preservation duration, facilitating ex vivo organ repair, and accelerating clinical translation.