We have demonstrated a direct correlation between a high ATP level in a canine pancreas graft after preservation by the two-layer method and good posttransplant outcome. The purpose of this study was to examine the effect of fasting and exogenous adenosine on the ATP tissue level and viability of the canine pancreas graft during preservation by the two-layer method. Graft viability was judged by graft survival following autotransplantation. Maintenance of normoglycemia for 5 days post-transplant was considered graft survival. The pancreas was harvested from either 72-hr-fated (n = 3) or -fed dogs (n = 4) and preserved by the two-layer (Euro-Collins' solution [EC]/perfluorochemical [PFC]) method for 24 hr. Graft survival rates in fed and fasted groups were 4/4 (100%) and 3/3 (100%), respectively. There was no significant difference in ATP tissue concentrations between the two groups (7.48 +/- 0.55 vs. 7.03 +/- 0.74 mumol/g dry wt, NS). The pancreas was subjected to 60 min warm ischemia and then was preserved by the two-layer method using EC or EC containing 5 mM adenosine for 24 hr. Without adenosine, graft survival rate was 0/3 (0%) and ATP tissue levels were not changed during preservation (1.62 +/- 0.26 vs. 1.56 +/- 0.40 mumol/g dry wt, NS). However, provision of adenosine to the graft during preservation led to the restoration of ATP tissue levels (1.90 +/- 0.54 vs. 8.13 +/- 0.98 mumol/g dry wt, P < 0.01) in 4 of 5 grafts, and these grafts functioned immediately and maintained normoglycemia after transplantation. Graft survival rate was 4/5 (80%). One of 5 grafts, however, did not survive, and the ATP tissue level was not adequately recovered during preservation compared with viable grafts (3.67 vs. 8.13 +/- 0.98 mumol/g dry wt). This study clearly demonstrates that the nutritional state of the donor has no influence on the ATP tissue level and viability of the graft during 24-hr preservation by the two-layer method. On the other hand, provision of adenosine to the graft during preservation stimulates ATP synthesis and improves the viability of the ischemically damaged pancreas.