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Optimizing Temperature and Oxygen Supports Long-term Culture of Human Islets

Komatsu, Hirotake and Rawson, Jeffrey and Medrano, Leonard and Cook, Colin A. and Barriga, Alyssa and Gonzalez, Nelson and Salgado, Mayra and Omori, Keiko and Kandeel, Fouad and Tai, Yu-Chong and Mullen, Yoko (2019) Optimizing Temperature and Oxygen Supports Long-term Culture of Human Islets. Transplantation, 103 (2). pp. 299-306. ISSN 0041-1337.

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Background: Islet transplantation is a promising treatment for type-1 diabetes; however, donor shortage is a concern. Even when a pancreas is available, low islet yield limits the success of transplantation. Islet culture enables pooling of multiple low-yield isolations into an effective islet mass, but isolated islets rapidly deteriorate under conventional culture conditions. Oxygen depletion in the islet core, which leads to central necrosis and volume loss, is one of the major reasons for this deterioration. Methods: To promote long-term culture of human islets in PIM-R medium (used for islet research), we adjusted temperature (12, 22, and 37°C) and oxygen concentration (21% and 50%). We simulated the oxygen distribution in islets based on islet oxygen consumption rate and dissolved oxygen in the medium. We determined the optimal conditions for oxygen distribution and volume maintenance in a 2-week culture and assessed viability and insulin secretion compared to noncultured islets. In vivo islet engraftment was assessed by transplantation into diabetic NOD-SCID mouse kidneys. We validated our results using CMRL 1066 medium (used for clinical islet transplantation). Results: Simulation revealed that 12°C–50% oxygen PIM-R culture supplied oxygen effectively into the islet core. This condition maintained islet volume at >90% for 2 weeks. There were no significant differences in viability and function in vitro or diabetic reversal rate in vivo between 2-week cultured and noncultured islets. Similar results were obtained using CMRL 1066. Conclusion: By optimizing temperature and oxygen concentration, we cultured human islets for 2 weeks with minimal loss of volume and function.

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Tai, Yu-Chong0000-0001-8529-106X
Additional Information:© 2018 Wolters Kluwer Health, Inc.
Record Number:CaltechAUTHORS:20180529-080857738
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Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:86645
Deposited By: Tony Diaz
Deposited On:29 May 2018 16:14
Last Modified:29 Jan 2019 17:47

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