Studies on islet cell viability atored with various preservatives : Assessment of adynylylcyclase activity as a marker for islet cell viability

对于临床胰岛细胞移植，可能需要短期的胰岛细胞滞留，在此期间胰岛细胞尽可能保持活力是至关重要的。然而，关于哪种保存液最适合于分离后或移植前的胰岛细胞保存的数据很少。我们注意到c-AMP是所有哺乳动物细胞中基本的第二信使，并使用新开发的荧光测定法在基础和刺激条件下测量了胰岛细胞c-AMP的产生，作为细胞活力的功能标记。Lewis大鼠胰岛分别保存不同时间（新鲜、3、16、24、48、96小时）和不同的保存溶液。UW （University of Wisconsin）溶液的腺苷酸环化酶活性分别为60.8(]SY.+-[)13.8、86.6(]SY.+-[)17.5、77.5(]SY.+-[)16.2、69.8(]SY.+-[)17.8、45.5(]SY.+-[)18.8和33.5(]SY.+- [)15.7 fmol/min/胰岛。刺激胰岛腺苷酸环化酶活性在欧-柯林斯（EC）和乳酸-林格（LR）溶液中保存3小时，分别为63.3(]SY.+- .[)20.3和53.3(]SY.+- . [)16.8 fmol/min/胰岛。在不同的保存时间内，UW溶液的腺苷酸环化酶活性显著高于EC和LR溶液。每个UW溶液保存组每只链脲佐菌素诱导的糖尿病大鼠移植约1000个胰岛细胞。通过测量血糖和体重来评估移植成功率。移植后3周，所有大鼠血糖均恢复正常。在糖尿病大鼠模型中，术前腺苷酸环化酶活性与移植后胰岛细胞功能密切相关。结果提示，胰岛保存时间（0、3、16、24小时）的差异不影响移植后胰岛细胞功能，术前腺苷酸环化酶活性与血浆葡萄糖恢复率有良好的相关性。进一步的研究表明，在人胰岛细胞中测量腺苷酸环化酶活性是可行的。我们得出结论，腺苷酸环化酶的活性可以作为胰岛细胞活力的有用标志，无论是实验还是临床。少

For clinical islet cell transplantation, short term sotrage of islet cell is likely to be necessary, and it is imperative that islet cell keeps to be as viable as possible during the period. However, there are little data on which preservative solutions are most suitable for the storage of islet cell after isolation or before transplantation.We noticed c-AMP is a fundamental second messenger in all mammalian cells, and measured c-AMP production of islet cell using a newly developed fluorometric assay under basal and stimulated conditions as a functional marker of cell viability.Islets prepared from Lewis rat were stored for different periods of time (fresh, 3,16,24,48,96 hours) and in different preservative solutions. The stimulated adenylylcyclase activities of UW (University of Wisconsin) solution were 60.8(]SY.+-。[)13.8,86.6(]SY.+-。[)17.5,77.5(]SY.+-。[)16.2,69.8(]SY.+-。[)17.8,45.5(]SY.+-。[)18.8, and 33.5(]SY.+-。[)15.7 fmol/min/islet, respectively. Stimulated adenylylcyclase activiti … More es of isolated islets stored with Euro-Collins (EC) and Lactate-Ringer (LR) solutions for 3 hours were 63.3(]SY.+-。[)20.3 and 53.3(]SY.+-。[)16.8 fmol/min/islet, respectively. Adenylylcyclase activity of UW solution was significantly higher than that of EC and LR solutions through the different periods of those preservation times.Approximately 1,000 islet cells in each preservation group of UW solution were transplanted per streptozotocin induced diabetes rat. Transplant success rate was evaluated by measuring blood glucose level and body weight. Three weeks after the transplant, all rats recovered euglycemia. Preoperative adenylylcyclase activity was correlated well with post-transplant islet cell function in a rat model of diabetes. The result suggests that differences in islet preservation time (0,3,16,24 hours) do not effect post-transplant islet cell function, and there is a good correlation between the preoperative adenylylcyclase activity and recovery rate of plasma glucose. Additional studies showed it was feasible to measure adenylylcyclase activity in human islet cells.We conclude that the activity of adenylylcyclase can be a useful marker of islet cell viability, both experimentally and clinically. Less