190 - 21°C is the New Normothermia for Organ Preservation in the Presence of H2S

Abstract

Background Cold storage (4°C) is the clinical standard of care for organ preservation prior to renal transplantation. However, it has been shown to cause negative graft outcomes. Normothermic alternatives are being investigated but there is no consensus on the ideal temperature. Organ preservation at physiological temperature (37°C) has shown some success but temperatures closer to room temperature could improve outcomes. Additionally, hydrogen sulfide (H2S) has been shown to mitigate renal injury during organ preservation. In this study, we investigate the effects of a novel, mitochondria-targeted H2S donor (AP39) using an in vitro model of normothermic preservation at 21°C. Methods Pig kidney epithelial cells (LLC-PK1) cells were treated with University of Wisconsin (UW) solution supplemented with 5nM, 50nM, 500nM or 5μM AP39. The cells were placed in hypoxia (0.5% O2) for 18h at 21°C, which simulates ischemia during normothermic preservation, followed by reoxygenation (21% O2) in Media 199 for 24h at 37°C. Cells were then stained with PE-Annexin-V and 7-AAD to quantify apoptosis and necrosis using flow cytometry. Results UW solution supplemented with 500nM AP39 and 5µM AP39 significantly increased (P Conclusion We demonstrate, for the first time, that normothermic preservation (21°C) with AP39-supplemented UW solution significantly increases renal cell viability and decreases renal cell apoptosis.