Kanazawa S, Tsunoda T, Murakami M, Iwamoto S, Kimoto M, Onuma E, Majima T, Yamamoto Y
Department of Surgery, Mitsugi General Hospital, Hiroshima, Mitsugi-gun, Japan.
Cell Transplant. 1999 Jul-Aug;8(4):383-8. doi: 10.1177/096368979900800406.
For clinical islet cell transplantation, short-term storage of islet cells is likely to be necessary, and it is imperative that the islet cells be kept as viable as possible during the period. However, there are little data on which preservative solutions are most suitable for the storage of islet cells after isolations or before transplantation. To estimate islet cell viability and transplantation success rate in the present study, adenylylcyclase activity was measured with a rapid new fluorometric assay in rat islet cells prior to transplantation, because cAMP plays an essential role in determining islet beta-cell viability and responsiveness to various hormonal stimuli. Adenylylcyclase activity was measured in islet cells stored for different periods of time 0, 3, 16, 24, 48, 96 h) and in different preservative solutions. Approximately 1,000 islet cells from each preservation group using University of Wisconsin (UW) solution were transplanted to streptozotocin-induced diabetes mellitus (DM) rats. Transplant success was evaluated by measuring blood glucose levels. Preoperative adenylylcyclase activity was compared with posttransplant islet cell function. The adenylylcyclase activity of UW solution was significantly higher than that of Euro-Collins solution and lactate-Ringer's solution through the different preservation time periods. Preoperative adenylylcyclase activity correlated well with posttransplant islet cell function in a rat model of DM. We conclude that adenylylcyclase activity can be used as a marker to assess islet cell viability as well as differences in preservation media and may predict islet cell transplant success.
对于临床胰岛细胞移植,胰岛细胞的短期储存可能是必要的,并且在此期间必须尽可能保持胰岛细胞的活力。然而,关于哪种保存液最适合胰岛细胞分离后或移植前的储存,相关数据很少。在本研究中,为了评估胰岛细胞活力和移植成功率,在大鼠胰岛细胞移植前,采用一种快速的新型荧光测定法测量腺苷酸环化酶活性,因为环磷酸腺苷(cAMP)在决定胰岛β细胞活力和对各种激素刺激的反应性方面起着至关重要的作用。在不同时间段(0、3、16、24、48、96小时)储存于不同保存液中的胰岛细胞中测量腺苷酸环化酶活性。使用威斯康星大学(UW)溶液将来自每个保存组的约1000个胰岛细胞移植到链脲佐菌素诱导的糖尿病(DM)大鼠体内。通过测量血糖水平评估移植成功率。将术前腺苷酸环化酶活性与移植后胰岛细胞功能进行比较。在不同的保存时间段内,UW溶液的腺苷酸环化酶活性显著高于欧洲柯林斯溶液和乳酸林格氏液。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。我们得出结论,腺苷酸环化酶活性可以用作评估胰岛细胞活力以及保存培养基差异的标志物,并且可能预测胰岛细胞移植的成功。