A simplified approach to human islet quality assessment.
A simplified approach to human islet quality assessment.
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DOI：
10.1097/tp.0b013e3181d54bce
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发表时间：
2010-05-27
期刊：
影响因子：
6.2
通讯作者：
Fernandez LA
中科院分区：
文献类型：
--
作者：
Hanson MS;Park EE;Sears ML;Greenwood KK;Danobeitia JS;Hullett DA;Fernandez LA
The successful treatment of patients with type 1 diabetes by islet transplantation is affected by a multitude of factors, of which infusion of the highest quality tissue is essential. The current standard pre-transplant quality assessments lack sensitivity, accuracy, and objectivity in the determination of islet viability and potency. We hypothesized that a multi-parametric approach focused on islet cell metabolic state, mitochondrial integrity, and in vitro glucose stimulated insulin secretion could provide data predictive of in vivo function. The objective of this study was to validate a novel set of islet quality assays and develop a simplified islet quality scoring system for both basic research and clinical applications. A series of 42 human islet preparations were screened using standard and novel methods, which included determination of yield, viability by fluorescent microscopy, glucose stimulated insulin secretion (GSIS), percentage of islet loss in culture, quantification of adenine nucleotides, flow cytometric measurement of viability, apoptosis, and mitochondrial membrane potential (MMP). In vivo functional potency was tested by minimal model transplant in streptozotocin-induced diabetic NOD.scid mice. Functionally potent islet preparations showed significantly greater numbers of cells with polarized MMP, higher ATP/ADP ratios and increased glucose induced insulin secretion. The MMP, ATP/ADP, and GSIS data were combined into a single islet scoring formula that showed >86% accuracy in predicting in vivo functional potency. Our study demonstrates that a multi-parametric approach using objective assessments focused on islet cell mitochondrial integrity and in vitro function can provide data predictive of in vivo function.
