THE POSSIBILITY OF ISLET ISOLATION FROM PANCREAS HARVESTED FROM NON-HEART BEATING CADAVER

Abstract

342 Introduction: Pancreas from non-heartbeating cadaver can be available in pancreas islet transplantation if its damage from warm ischamia (WI) could be reduced by a suitable method. The two-layer cold storage method (TL), which utilizes perfluorochemical and UW solution, has previously demonstrated to have unique characteristics of supplying oxygen to the graft, effectively maintaining the viability of whole pancreas grafts. In this study we evaluated the effect of TL on islet yield and viability of WI damaged pancreas. Method: Dog pancreases were divided into following five groups (group 1; n=10, groups 2∼5; n=5). In group 1, pancreases were processed immediately after total pancreatectomy. In groups 2 (4), pancreases were left in the abdominal cavity for 60 (90) minutes as WI, then processed into islets. In groups 3 (5), pancreases were left in the abdominal cavity for 60 (90) minutes as WI followed by TL for 24 hours at 4°C, then processed into islets. Islet isolation was performed by the automated collagenase digestion method of Ricordi, purified on Euro-Ficoll discontinuous gradients. We compared five groups in terms of islet yield, expressed as islet equivalents (IEQ, diameter standardizing to 150 μm) per pancreas weight (IEQ/g pancreas). Results: Islet yield was 2700 ± 1200 (mean±SD) in group 1, 1500 ± 700 in group 2, (p=0.03 vs. group 1, p=0.02 vs. group 3), and 2600 ± 600 in group 3 (p=0.80 vs. group 1). In group 4 and 5, almost no islets were obtained after purification. In static incubation islets in group 1 and 3 showed sufficient insulin release in response to glucose stimuli and this means that viability of islets in group 1 and 3 was maintained. Conclusions: These data clearly demonstrates that TL following WI can regain islet yield and viability almost at the same level as those of fresh pancreas. So it could be possible to isolate islets from pancreas harvested from non-heartbeating cadaver by TL.