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Chronic shortage of donor organs has led to acceptance of steatotic livers as grafts, although there is a higher risk of primary graft dysfunction. We herein report the beneficial impact of Polysol, a newly developed preservation solution, on cold storage of steatotic rat livers. Dietary hepatic steatosis was induced in Wistar rats by 2-day fasting and subsequent 3-day re-feeding with a fat-free, carbohydrate-rich diet. Fatty livers were retrieved, flushed and then stored at 4°C for 24 hours with either HTK or Polysol. Functional integrity of the grafts was evaluated by isolated reperfusion with oxygenated Krebs-Henseleit buffer at 37°C for 45 minutes in both groups. Polysol preservation resulted in significant reductions of not only parenchymal (AST (IU/L); 6728±824 in HTK vs. 3107±718 in Polysol; P < 0.001) but also mitochondrial (GLDH (IU/L); 3189±773 vs. 1282±365; P < 0.01) enzyme release throughout reperfusion. Moreover, PVP (16.9±2.7 vs. 7.8±1.5 mmHg; P < 0.05), hepatic O2 consumption (0.291±0.047 vs. 1.056±0.053 μmol/g liver/min; P < 0.001), tissue ATP content (0.695±0.086 vs. 1.340±0.157 μmol/g dry-liver; P < 0.005), bile production (0.79±0.11 vs. 4.08±0.66 μL/g liver/45-min; P < 0.001), malondialdehyde into the perfusate (1.922±0.198 vs. 0.573±0.094 nmol/L; P < 0.0001) and wet/dry-weight ratio of the liver tissues (5.20±0.31 vs. 3.85±0.15; P < 0.005) were all better preserved by Polysol. In line with these benefits, electron microscopy revealed that Polysol preservation substantially suppressed deleterious mitochondrial alterations in steatotic livers. In conclusion, cold storage using Polysol resulted in significantly better integrity and function of steatotic livers. Polysol, therefore, may be a new alternative especially for “marginal” organs. Liver Transpl, 2006. © 2006 AASLD.