DEVELOPMENT OF METHODS TO IMPROVE ISOLATED PANCREATIC ISLET FUNCTION

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 目的：开发新的分离和评价分离胰岛的方法，以移植到I型糖尿病患者中。 使用非人灵长类动物组织获得的结果将有助于开发用于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，作为移植前胰岛制剂质量的快速效力指标。2)测定分离后缺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细胞凋亡的敏感性。 将使用一种新方法进行活力和凋亡的测定，其中可以使用复杂对象参数分析仪和分选仪（COPAS）流式细胞术完整分析胰岛，保留其完整结构。我们还将评估COPAS活力和细胞凋亡测量的能力，以提供移植后胰岛功能的预测数据，在链脲佐菌素诱导的糖尿病免疫缺陷小鼠中使用人胰岛移植的边缘质量模型。此外，开发快速、准确和预测性的胰岛活性和分离后功能测试将有助于显著改善胰岛功能。 改善移植后的成功率。这项研究使用WNPRC研究服务。 注意事项：在本报告期内，该研究者没有任何涉及灵长类动物中心的项目、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，我们将该子项目再纳入一年，以证明他最近的非人灵长类动物工作与他目前在该领域的人类研究之间的重要联系。 出版物待定。

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.