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

用各种防腐剂对胰岛细胞活力的研究：评估作为胰岛细胞活力标记的腺苷酰环化酶活性

• 批准号: 07457283
• 负责人: TSUNODA Tsukasa
• 金额: $ 3.39万
• 依托单位: KAWASAKI MEDICAL SCHOOL
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07457283/
• 关键词: islet cell transplantation; c-AMP; adenylylcyclase activity; diabetes mellitus; islet cell viability; UW solution; total pancreatectomy; autotransplantation; diabees mellitus; C-AMP; enzymatic cycling; adenylate cyclase activity; Hanks solutio

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

临床胰岛细胞移植需要短期的胰岛细胞复苏，在此期间尽可能保持胰岛细胞的活力是非常必要的。然而，目前关于胰岛细胞分离后或移植前的保存液最合适的数据很少。我们注意到c-AMP是所有哺乳动物细胞中最基本的第二信使，测量C-应用荧光法测定基础和刺激条件下胰岛细胞AMP的生成量，作为细胞活力的功能性指标时间（新鲜，3，16，24，48，96小时）和不同的防腐剂溶液。UW（University of威斯康星州）溶液刺激的腺苷酸环化酶活性为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. ±. [）15.7fmol/min/胰岛。腺苷酸环化酶激活 ...更多信息 用Euro-Collins（EC）和Lactate-Ringer（LR）溶液保存3小时的胰岛细胞的ES为63.3（± SY）。[）20.3和53.3（]SY. ±. [）16.8fmol/min/胰岛。UW液保存组的腺苷酸环化酶活性在不同时间段均显著高于EC和LR液。通过测量血糖水平和体重来评估移植成功率。移植后三周，所有大鼠均恢复正常。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 h）的不同对移植后胰岛细胞功能无影响，术前腺苷酸环化酶活性与血糖恢复率有良好的相关性。进一步的研究表明，腺苷酸环化酶活性的测定在人类胰岛细胞中是可行的。我们的结论是，腺苷酸环化酶的活性可以是一个有用的标记胰岛细胞的活力，无论是实验和临床。少

项目成果

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T.Tsunoda、Y.Yamamoto、M.Kimoto、H.Imai、S.Iwamoto、S.Kawasaki、K.Kawashima、Y.Tadaoka、T.Majima、E.Onuma、K.Iki、T.Kubozoe、T.

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