ISLET ISOLATION ASSESSMENT IN MAN AND LARGE ANIMALS
ISLET ISOLATION ASSESSMENT IN MAN AND LARGE ANIMALS
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DOI：
10.1007/bf02581331
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发表时间：
1990-07-01
期刊：
影响因子：
--
通讯作者：
LACY, PE
中科院分区：
文献类型：
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作者：
RICORDI, C;GRAY, DWR;LACY, PE
从包括人在内的大型哺乳动物的胰腺中分离胰岛的最新进展，强调需要发展精确和可重复的技术来评估胰岛产量。本文讨论了胰岛分离评估的定量和定性标准，主要内容是最终胰岛制剂的数量、体积、纯度、形态完整性和体外和体内功能测试的确定。推荐双硫腙作为一种特异性染色剂，用于即时检测胰岛组织，从而可以估计胰岛的总数（将其分为50µ直径范围增量的类别）和最终制剂的纯度。适当的形态学评估应包括确认胰岛鉴定，评估形态学完整性和胰岛制剂的纯度。使用荧光包合和排除染料一起被建议作为一种活力测定，同时定量细胞的比例是完整的或受损的。胰岛的葡萄糖浸润提供了葡萄糖介导的胰岛素释放的动态概况，以及细胞在血糖挑战中断后下调胰岛素分泌的能力。虽然灌注数据为胰岛活力提供了有用的指导，但胰岛素释放的数量和动力学并不一定能预测植入后胰岛的性能。因此，胰岛活力的最终测试是它们移植到糖尿病受体后的功能。因此，有人建议将最终胰岛制剂的等量移植到糖尿病裸鼠（胸腺）体内。我们希望这些一般准则将有助于标准化胰岛隔离的评估，从而能够更好地解释和比较不同中心的程序。
Recent progress in islet isolation from the pancreas of large mammals including man, accentuated the need for the development of precise and reproducible techniques to assess islet yield. In this report both quantitative and qualitative criteria for islet isolation assessment were discussed, the main topics being the determination of number, volume, purity, morphologic integrity andin vitroandin vivofunction tests of the final islet preparations. It has been recommended that dithizone should be used as a specific stain for immediate detection of islet tissue making it possible to estimate both the total number of islets (dividing them into classes of 50 µ diameter range increments) and the purity of the final preparation. Appropriate morphological assessment should include confirmation of islet identification, assessment of the morphological integrity and of the purity of the islet preparation. The use of fluorometric inclusion and exclusion dyes together have been suggested as a viability assay to simultaneously quantitate the proportion of cells that are intact or damaged. Perifusion of islets with glucose provides a dynamic profile of glucose-mediated insulin release and of the ability of the cells to down regulate insulin secretion after the glycemic challenge is interrupted. Although perifusion data provides a useful guide to islet viability the quantity and kinetics of insulin release do not necessarily predict islet performance after implantation. Therefore, the ultimate test of islet viability is their function after transplantation into a diabetic recipient. For this reason,in vivomodels of transplantation of an aliquot of the final islet preparation into diabetic nude (athymic) rodents have been suggested. We hope that these general guidelines will be of assistance to standardize the assessment of islet isolations, making it possible to better interpret and compare procedures from different centers.