This subproject is one of many research subprojects utilizing theresources provided by a Center grant funded by NIH/NCRR. The subproject andinvestigator (PI) may have received primary funding from another NIH source,and thus could be represented in other CRISP entries. The institution listed isfor the Center, which is not necessarily the institution for the investigator.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.Dr. Fernandez also has three pending publications related to his nonhuman primate work and we will include those in next year's APR.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.
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