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用于评估人胰岛功能的自动化高通量检测方法

Automated, High-Throughput Assays for Evaluation of Human Pancreatic Islet Function.

作者信息

Cabrera Over, Jacques-Silva M Caroline, Berman Dora M, Fachado Alberto, Echeverri Felipe, Poo Ramon, Khan Aisha, Kenyon Norma S, Ricordi Camillo, Berggren Per-Olof, Caicedo Alejandro

机构信息

Diabetes Research Institute, Miller School of Medicine, University of Miami, Miami, FL, USA.

The Rolf Luft Research Center for Diabetes and Endocrinology, Karolinska Institutet, Stockholm, Sweden.

出版信息

Cell Transplant. 2007 Nov;16(10):1039-1048. doi: 10.3727/000000007783472408.

Abstract

An important challenge in pancreatic islet transplantation in association with type 1 diabetes is to define automatic high-throughput assays for evaluation of human islet function. The physiological techniques presently used are amenable to small-scale experimental samples and produce descriptive results. The postgenomic era provides an opportunity to analyze biological processes on a larger scale, but the transition to high-throughput technologies is still a challenge. As a first step to implement high-throughput assays for the study of human islet function, we have developed two methodologies: multiple automated perifusion to determine islet hormone secretion and high-throughput kinetic imaging to examine islet cellular responses. Both technologies use fully automated devices that allow performing simultaneous experiments on multiple islet preparations. Our results illustrate that these technologies can be applied to study the functional status and explore the pharmacological profiles of islet cells. These methodologies will enable functional characterization of human islet preparations before transplantation and thereby provide the basis for the establishment of predictive tests for β-cell potency.

摘要

1型糖尿病相关的胰岛移植中的一个重要挑战是定义用于评估人胰岛功能的自动高通量检测方法。目前使用的生理学技术适用于小规模实验样本,并产生描述性结果。后基因组时代为大规模分析生物过程提供了机会,但向高通量技术的转变仍然是一个挑战。作为实施用于研究人胰岛功能的高通量检测方法的第一步,我们开发了两种方法:用于确定胰岛激素分泌的多次自动灌流法和用于检查胰岛细胞反应的高通量动力学成像法。这两种技术都使用全自动设备,允许对多个胰岛制剂同时进行实验。我们的结果表明,这些技术可用于研究胰岛细胞的功能状态并探索其药理学特征。这些方法将能够在移植前对人胰岛制剂进行功能表征,从而为建立β细胞潜能的预测性检测提供基础。

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