AUTOMATED-METHOD FOR ISOLATION OF HUMAN PANCREATIC-ISLETS
AUTOMATED-METHOD FOR ISOLATION OF HUMAN PANCREATIC-ISLETS
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DOI：
10.2337/diabetes.37.4.413
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发表时间：
1988-04-01
期刊：
影响因子：
7.7
通讯作者：
SCHARP, DW
中科院分区：
文献类型：
--
作者：
RICORDI, C;LACY, PE;SCHARP, DW
我们描述了一种分离人类胰岛的自动化方法。该程序满足以下要求：1）对胰岛的创伤最小，2）连续消化，其中逐渐释放的胰岛可以免于进一步的酶作用，3）消化过程中最小的人为干预，以及4）分离胰岛的高产量和纯度。经过Ficoll梯度纯化后，平均获得164,600个胰岛/胰腺（2279个胰岛/g），平均胰岛纯度为78.5%。最终胰岛制剂的平均体积和平均胰岛素含量分别为 348 mm3 和 93.4 U。胰岛形态完整，具有正常程度的β-颗粒化，并且对葡萄糖刺激作出反应，胰岛素分泌比基础水平增加五倍。该程序目前正用于启动人类胰岛移植的第二阶段临床试验。
We describe an automated method for the isolation of human pancreatic islets. The procedure meets the following requirements: 1) minimal traumatic action on the islets, 2) continuous digestion in which the islets that are progressively liberated can be saved from further enzymatic action, 3) minimal human intervention in the digestion process, and 4) high yield and purity of the isolated islets. After purification on Ficoll gradients, an average of 164,600 islets/pancreas was obtained (2279 islets/g), with an average purity of 78.5% islets. The average volume and average insulin content of the final islet preparation were 348 mm3 and 93.4 U, respectively. The islets were morphologically intact with a normal degree of .beta.-granulation and responded to glucose stimulation with a fivefold increase of insulin secretion over basal levels. The procedure is now being used for the initiation of the second phase of clinical trials on human islet transplants.