Development of Apparatus for the Vitrification of Curative Doses of Human Islets for Transplantation

Project Summary Abstract The ultimate objective of the proposed research is to enable the long term banking of curative doses of human pancreatic islets for the reversal of typediabetes and the amelioration of typediabetesBanking should reduce or prevent the deterioration of islets prior to usefacilitate tolerance inductionenable single rather than multiple islet infusions to achieve remission of typediabetesenable use of many more islets for therapeutic or research purposesand have other advantagesWe have already demonstratedin unpublished resultsthat it is possible to vitrify and rewarm human pancreatic islets with excellent islet number recovery and excellent retention of viabilitybased on vital stainingglucose stimulated insulin releasestimulation indexglucose stimulated oxygen consumptionand ability to reverse induced diabetes in mice with minimal islet dosesThe goal of the proposed research is to demonstrate that it is possible to scale up from the islet numbers vitrified to dateup toIEQsto curative doses of isletswith no significant changes in cryoprotection techniquecooling rateor warming rateThis goal will be achieved by improving the spatial efficiency of the apparatus already created while simultaneously increasing the carrying capacity of that apparatusAimmilestoneis devoted to optimizing the balance between the rate of introduction of cryoprotective agents and the behavior of the islets within one specific and nominally optimal device designAimmilestonewill be devoted to exploring the design space of the device based on empirical observations of the responses of islets to different carrying capacity modelsIn Aimwe will fabricate and preliminarily test the optimized device resulting from the information gained in AimThis device will be mostly automated to enable use by relatively unskilled personnel and to prevent variations caused by operator factors or operator errorIn Aimwe will test the optimized device and method by actually vitrifying islets in densities within the device equivalent to the densities that will be required for preserving fully curative clinical islet dosesand the results will be evaluated independently by the City of Hope using the abovedescribed endpointsFinallyin Aimwe will obtain an objective evaluation of the barriers to clinical use of our methods and device as prepared by the City of Hopeand we will prepare a plan for overcoming any remaining barriers to clinical application based on the City of Hope reportThis should enable us to move forward toward clinical trials of vitrified curative doses of human pancreatic islets Project Narrative The ultimate objective of this application is to enable the cure of typediabetesT Dby making it possible to aprovide pancreatic islets to T D patients at optimal times and with minimal islet deterioration prior to transplantationbprovide tissue engineered substitutes for pancreatic islets when they become available by removing inventory control obstaclesthereby enabling all T D victims to receive curative insulin producing cellsand cinduce immunological tolerance to islet transplantsthereby overcoming both ordinary rejection and autoimmune rejectionThe common denominator between all of these requirements for curing T D is the ability to bank islets in excellent condition as long as may be required to aenable their transplantation into recipients under ideal conditionsbeliminate mismatches between the moment to moment supply and demand of laboratory produced islet replacementsand cenable tolerance to be induced prior to or simultaneously with the transplantation of donor or laboratory produced islets or islet equivalentsOur laboratory has already demonstrated the ability to bank human islets with excellent number recoveryviabilityglucose stimulated insulin releaseglucose stimulated oxygen consumption responsesand ability to cure induced T D in mice with the subcapsular transplantation of minimal islet dosesand thereforethe purpose of the present application is simply to scale up our existing successful methods to enable the vitrification of curative doses of human isletswhich appears to be quite possible based on preliminary calculations