DEVELOPMENT OF METHODS TO IMPROVE ISOLATED PANCREATIC ISLET FUNCTION

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 目的：建立分离和鉴定胰岛的新方法，为I型糖尿病患者的胰岛移植奠定基础。 利用非人灵长类组织获得的结果将有助于开发分离和评估人类胰岛的方法。方法包括：1)表征对葡萄糖的生理反应，作为移植前胰岛制备质量的快速效力指示器；2)分离后测定低氧对胰岛活力和功能的影响；3)测定胰岛对细胞因子诱导的凋亡的敏感性。将使用一种新的方法来确定存活和凋亡，在这种方法中，可以完整地分析胰岛，使用复杂对象参数分析仪和分类器(COPAS)流式细胞术保留其完整的结构。我们还将使用链脲佐菌素诱导的糖尿病免疫缺陷小鼠的人胰岛移植边缘质量模型，评估COPAS活性和细胞凋亡测量为胰岛移植后功能提供预测数据的能力。此外，分离后胰岛活性和功能的快速、准确和预测性测试的发展将有助于显著 移植后成功率的提高。这项研究使用了WNPRC Research Services。 注：在本报告所述期间，这位研究人员没有任何涉及灵长类中心的项目、资金或出版物；但他积极参与与这项工作相关的临床研究、资金和出版物；因此，我们将这个子项目再增加一年，以证明他最近在这一领域的非人类灵长类动物工作与他目前的人类研究之间的重要联系。 出版物正在等待发布。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Objective: To develop new methods for the isolation and evaluation of isolated pancreatic islets for the purpose of transplantation into patients with type I diabetes. The results obtained using non-human primate tissue will assist in the development of methodologies for the isolation and assessment of human islets. Methods include; 1) characterization of the physiological response to glucose as a rapid potency indicator of the quality of the islet preparation prior to transplantation,. 2) determination of the effects of hypoxia on islet viability and function after isolation, 3) determination of the sensitivity of islets to cytokine induced apoptosis. Determination of viability and apoptosis will be performed using a novel approach in which islets can be analyzed intact, preserving their complete architecture using Complex Object Parametric Analyzer and Sorter (COPAS) flow cytometry. We will also evaluate the ability of COPAS viability and apoptosis measurements to provide predictive data on islet function after transplantation using a marginal mass model of human islet transplant in streptozotocin-induced diabetic immuno-deficient mice. In addition, the development of rapid, accurate, and predictive tests of islet viability and function post isolation will contribute to significant improvements in post transplant success. This research used WNPRC Research Services. NOTE: This investigator did not have any projects, funding or publications involving the Primate Center during this reporting period; but he is active with clinical research, funding and publications related to this work; thus, we are including this subproject for an additional year to demonstrate the important link between his recent nonhuman primate work and his current human research in this area. Publications are pending.