Comparison of Human Pancreatic Islet Viability & Functional Potency Following Culture & Mock Shipment in Silicone Rubber Membrane Bottles
Funding:
Beckman Research Institute / National Institutes of Health
Principal Investigator:
Lab Website:
Project Summary:
Standard protocols of islet culture and shipment are critical to successfully meeting the Islet Cell Resource program’s core mission of distributing human islets of the highest quality to researchers world-wide. In addition, islet culture pre-transplant has become the standard practice for clinical islet transplantation programs to facilitate quality control assessment and recipient patient pre-infusion management. Because islets are a multi-cellular aggregate with unique nutrient and oxygen demands, in vitro culture systems must be carefully designed to meet these needs. Moreover, the large quantity of islets that result from a successful human isolation (~250,000 islet particles/500,000 islet equivalents) complicate experimentation and distribution. Shipment of human islets from their laboratory of isolation to distal research centers has been shown to decrease viability and functional potency. Together, these problems are the impetus for studies to improve the quality of human islets after extended periods of culture or shipping. Tissue culture and shipping bottles with gas permeable silicone rubber membrane (SRM) bottoms represent a potential remedy for solving both the problems of islet culture seeding densities and proper nutrient and gas suppy. Increasing oxygenation of the culture media also allows for increased islet seeding densities, which decreases the required number of culture vessels by more than 10 fold. Evaluation of the viability and functional potency of human islets cultured and stored in SRM bottles will be performed in comparison to contol islets cultured and stored in standard flasks. We will assess islet quality using novel methods of measuring mitochondrial health and by transplantation into diabetic immunodeficient mice. Validation of a standardized islet culture and shipping methodology will improve the quality of islets distributed to researchers and potentially improve the post transplant outcomes in patients receiving islets cultured under conditions that are more favorable for promotion of viability and functional potency.
Type I diabetes continues to take a tremendous tole on citizens of the United States with over 1 million suffering from the disease and billions spent for medical diagnosis and treatment. Islet transplantation represents a successful therapeutic remedy for this disease that can be expanded in its accessibility to patients as improvements in the methods of islet isolation, culture, and transplantation are made.
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First published: 07/15/02 Last updated: 11/24/09
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