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Normothermic machine perfusion (NMP) provides clinicians an opportunity to assess marginal livers before transplantation. However, objective criteria and point‐of‐care (POC) biomarkers to predict risk and guide decision making are lacking. In this investigation, we characterized trends in POC biomarkers during NMP and compared primate donation after circulatory death (DCD) livers with short and prolonged warm ischemic injury. Following asystole, livers were subjected to either 5 minutes (DCD‐5min, n = 4) or 45 minutes (DCD‐45min, n = 4) of warm ischemia time. Livers were flushed with heparinized UW solution, and preserved in cold storage before NMP. During flow‐controlled NMP, circulating perfusate and tissue biopsies were collected at 0, 2, 4, 6, and 8 hours for analysis. DCD‐45min livers had greater terminal portal vein pressure (8.5 vs. 13.3 mm Hg,  P  = 0.027) and terminal portal vein resistance (16.3 vs. 32.4 Wood units,  P  = 0.005). During perfusion, DCD‐45min livers had equivalent terminal lactate clearance (93% vs. 96%,  P  = 0.344), greater terminal alanine aminotransferase (163 vs. 883 U/L,  P  = 0.002), and greater terminal perfusate gamma glutamyltransferase (GGT) (5.0 vs. 31.7 U/L,  P  = 0.002). DCD‐45min livers had higher circulating levels of flavin mononucleotide (FMN) at hours 2 and 4 of perfusion (136 vs. 250 ng/mL,  P  = 0.029; and 158 vs. 293 ng/mL,  P  = 0.003; respectively). DCD‐5min livers produced more bile and demonstrated progressive decline in bile lactate dehydrogenase, whereas DCD‐45min livers did not. On blinded histologic evaluation, DCD‐45min livers demonstrated greater injury and necrosis at late stages of perfusion, indicative of nonviability.  Conclusion:  Objective criteria are needed to define graft viability during NMP. Perfusate lactate clearance does not discriminate between viable and nonviable livers during NMP. Perfusate GGT and FMN may represent POC biomarkers predictive of liver injury during NMP.

Point‐of‐Care Assessment of DCD Livers During Normothermic Machine Perfusion in a Nonhuman Primate Model