Effects of cryopreservation on in vitro and in vivo long-term function of human islets
Effects of cryopreservation on in vitro and in vivo long-term function of human islets
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DOI：
10.1097/00007890-199909150-00011
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发表时间：
1999-09-15
期刊：
影响因子：
6.2
通讯作者：
Di Carlo, V
中科院分区：
文献类型：
--
作者：
Piemonti, L;Bertuzzi, F;Di Carlo, V
背景。移植后几天进行移植的可能性似乎是胰岛移植的一个特点，并且冷冻保存人类胰岛的机会可能允许一个无限期的时间来调节受体免疫系统。本研究的目的是评价冷冻保存的人胰岛的体外和体内功能特性。我们连续使用了六种人类胰岛制剂，由于有限的胰岛块分离，不适合糖尿病患者的立即移植。通过测定胰岛素和胰高血糖素在葡萄糖（16.7 mM）、葡萄糖(16.7 mM) + 3-异丁基-1-甲基黄嘌呤（0.1 mM）、精氨酸（10 mM）和甲苯丁酰胺（100 μ M）等经典刺激下的体外功能。在移植1000个人工挑选的新鲜或低温保存的裸鼠胰岛后15、30、60和90天，通过静脉葡萄糖耐量试验评估体内胰岛功能。冷冻胰岛的真胰岛素分泌量明显高于新鲜胰岛。冷冻保存的胰岛对精氨酸和葡萄糖+异丁基甲基黄嘌呤的反应似乎部分受损。对于所有使用的促分泌剂，冷冻保存的胰岛胰岛素样原分子分泌似乎高于新鲜胰岛，精氨酸的差异有统计学意义。人体冷冻胰岛维持糖尿病裸鼠正确代谢控制的能力在3个月内逐渐丧失。这些发现表明，低温保存影响离体胰岛的功能，在有限的时间内维持体内功能。
Background. The possibility of performing transplantation several days after 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 evaluation of in vitro and in vivo functional properties of cryopreserved human islets.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 classical stimuli as glucose (16.7 mM), glucose (16.7 mM) + 3-isobutyl-1-methylxanthine (0.1 mM), arginine (10 mM), and tolbutamide (100 mu M). In vivo isfet function was assessed through intravenous glucose tolerance tests performed 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 cryopreserved 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 progressively lost in 3 months.Conclusions. These findings showed that cryopreservation affects the function of isolated human islets, maintaining in vivo function for a limited period of time.