Effect of different warm ischemic time on islet isolation and function in pigs

Abstract

Objective To study the effect of different warm-ischemia time on islet structure, yield and function. Methods Adult pigs were studied, and according to warm ischemic time (WIT, 0, 10, 20, 30, 45 min), the porcine pancreata were divided into 5 groups (A, B, C, D, and E group). The isolation of adult porcine pancreas was carried out by injection of Hanks into pancreatic duct and digestion with collagenase Ⅴ and Ⅰ. A discontinuous Ficoll 400 solution was applied for purifying islets. The structure, purity and viability of islets were determined by dithizone staining and AO/PI staining, and the function of islets was evaluated by the glucose stimulating insulin release test. Results There were little islets debris in group A, B and C. But it was very difficult to obtain integrate islets in group E. There was no significant difference between the groups of 0, 10 and 20 min in islet number before and after islet purification ( P ＞ 0.05 ), while the difference was significant when warm ischemic time was prolonged to 30 min and 45 min ( P ＜ 0. 01 ). In respect of islet function, the similar results were found in different groups. Conclusions When WIT is within 20 min, there is no deleterious effects on islet morphology and yield as well as function. However, when WIT is prolonged to 30 min, the loss in islet yield is severe, and the islet function is significantly impacted. Key words: Adult pig; Islet; Warm ischemia