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人胰岛分离的自动化方法。

Automated method for isolation of human pancreatic islets.

作者信息

Ricordi C, Lacy P E, Finke E H, Olack B J, Scharp D W

机构信息

Department of Pathology, Washington University School of Medicine, St. Louis, MO 63110.

出版信息

Diabetes. 1988 Apr;37(4):413-20. doi: 10.2337/diab.37.4.413.

Abstract

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.

摘要

我们描述了一种用于分离人胰岛的自动化方法。该程序满足以下要求:1)对胰岛的创伤作用最小;2)连续消化,在此过程中逐步释放的胰岛可免受进一步的酶作用;3)消化过程中的人工干预最少;4)分离出的胰岛产量高且纯度高。在Ficoll梯度上纯化后,平均每个胰腺获得164,600个胰岛(2279个胰岛/克),胰岛平均纯度为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立方毫米和93.4单位。胰岛形态完整,β颗粒化程度正常,对葡萄糖刺激的反应是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胰岛移植临床试验的第二阶段。

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