Effects of cryopreservation on in vitro and in vivo long-term function of human islets

Background. The possibility of performing transplantation several days afte r explant seems to be a peculiarity of islet grafts, and the opportunity to cryo-preserve human islets may permit an indefinite period for modulating the recipient immune system. The aim of the present study was the evaluatio n of in vitro and in vivo functional properties of cryopreserved human isle ts. Methods. We used six consecutive human islet preparations not suitable for an immediate transplantation in diabetic patients because the limited islet mass separated. The in vitro function of cryo and fresh islets was studied by determination of insulin and glucagon secretion in response to such cla ssical stimuli as glucose (16.7 mM), glucose (16.7 mM) + 3-isobutyl-1-methy lxanthine (0.1 mM), arginine (10 mM), and tolbutamide (100 mu M). In vivo i sfet function was assessed through intravenous glucose tolerance tests perf ormed at 15, 30, 60, and 90 days after transplantation of 1000 hand-picked fresh or cryopreserved islets in nude mice. Results. Basal secretion of true insulin was significantly higher in cryopr eserved islets than in fresh ones. The response of cryopreserved islets to arginine and glucose + isobutyl-l-methylxanthine seemed partially impaired. Proinsulin-like molecule secretion seemed higher in cryopreserved than in fresh islets in response to all secretagogues used, and the difference was statistically significant for arginine. The capacity of human cryopreserved islets to maintain a correct metabolic control in diabetic nude mice was p rogressively lost in 3 months. Conclusions. These findings showed that cryopreservation affects the functi on of isolated human islets, maintaining in vivo function for a limited per iod of time.