Normothermic ex vivo kidney perfusion (NEVKP) represents a novel approach for graft preservation and functional improvement in kidney transplantation. We investigated whether NEVKP also allows graft quality assessment before transplantation. Kidneys from 30-kg pigs were recovered in a model of heart-beating donation (group A) after 30minutes (group B) or 60minutes (group C) (n=5/group) of warm ischemia. After 8hours of NEVKP, contralateral kidneys were resected, grafts were autotransplanted, and the pigs were followed for 3days. After transplantation, renal function measured based on peak serum creatinine differed significantly among groups (P<.05). Throughout NEVKP, intrarenal resistance was lowest in group A and highest in group C (P<.05). intrarenal resistance at the initiation of NEVKP correlated with postoperative renal function (P<.001 at NEVKP hour 1). Markers of acid-base homeostasis (pH, HCO3- , base excess) differed among groups (P<.05) and correlated with posttransplantation renal function (P<.001 for pH at NEVKP hour 1). Similarly, lactate and aspartate aminotransferase were lowest in noninjured grafts versus donation after circulatory death kidneys (P<.05) and correlated with posttransplantation kidney function (P<.001 for lactate at NEVKP hour 1). In conclusion, assessment of perfusion characteristics and clinically available perfusate biomarkers during NEVKP allows the prediction of posttransplantation graft function. Thus, NEVKP might allow decision-making regarding whether grafts are suitable for transplantation.