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A COMPARISON OF FOUR SOLUTIONS FOR COLD STORAGE OF PANCREATIC ISLETS1

Vinícius Daher Alvares Delfino, DEREK W. R. GRAY, C. Kar Leow, Shinichi Shimizu, D. Ferguson, Peter J. Morris

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Abstract

Clinical pancreatic islet transplantation requires cold storage of islets for some hours. The best solution for cold storage will be that which maintains islet viability for the longest time. We have examined the viability of rat and human islet after storage at 4 degrees C for up to 6 days in Hanks' balanced salt solution, University of Wisconsin solution (UW), and 2 modified versions of this last solution, Sumimoto D (SD) and histidine-lactobionate (HL). The integrity of cold-stored rat and human islets of Langerhans has been examined using supravital stains and electron microscopy. In addition, the viability of cold-stored rat islets was tested by intraportal transplantation into syngeneic streptozotocin-induced diabetic recipients. The in vitro studies showed good preservation of islets stored in UW, SD, and HL for up to 72 hr. In comparison, storage for periods as short as 24 hr in HBSS markedly reduced islet integrity. The transplantation studies showed that rat islets cold stored in HBSS solution for 24 hr were not able to reverse experimental diabetes, whereas those stored in the other 3 solutions for 24 hr successfully reversed diabetes within 1 week of transplantation. After 48-hr cold storage, only islets preserved in HL solution were uniformly capable of producing functioning islet grafts. None of the tested solutions was able to maintain islet viability sufficiently to allow successful transplantation after more than 48 hr of cold storage. These experiments demonstrate that good islet viability is maintained for up to 24 hr of storage in UW, SD, and HL, and even after 48-hr cold storage in HL solution, whereas preservation in HBSS solution was deleterious to islet viability within 24 hr.

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What this paper is about

Clinical pancreatic islet transplantation requires cold storage of islets for some hours. The best solution for cold storage will be that which maintains islet viability for the longest time. We have examined the viability of rat and human islet after storage at 4 degrees C for up to 6 days in Hanks' balanced salt solution, University of Wisconsin solution (UW), and 2 modified versions of this last solution, Sumimoto D (SD) and histidine-lactobionate (HL). The integrity of cold-stored rat and human islets of Langerhans has been examined using supravital stains and electron microscopy. In addition, the viability of cold-stored rat islets was tested by intraportal transplantation into syngeneic streptozotocin-induced diabetic recipients. The in vitro studies showed good preservation of islets stored in UW, SD, and HL for up to 72 hr. In comparison, storage for periods as short as 24 hr in HBSS markedly reduced islet integrity. The transplantation studies showed that rat islets cold stored in HBSS solution for 24 hr were not able to reverse experimental diabetes, whereas those stored in the other 3 solutions for 24 hr successfully reversed diabetes within 1 week of transplantation. After 48-hr cold storage, only islets preserved in HL solution were uniformly capable of producing functioning islet grafts. None of the tested solutions was able to maintain islet viability sufficiently to allow successful transplantation after more than 48 hr of cold storage. These experiments demonstrate that good islet viability is maintained for up to 24 hr of storage in UW, SD, and HL, and even after 48-hr cold storage in HL solution, whereas preservation in HBSS solution was deleterious to islet viability within 24 hr.

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Available abstract

Clinical pancreatic islet transplantation requires cold storage of islets for some hours. The best solution for cold storage will be that which maintains islet viability for the longest time. We have examined the viability of rat and human islet after storage at 4 degrees C for up to 6 days in Hanks' balanced salt solution, University of Wisconsin solution (UW), and 2 modified versions of this last solution, Sumimoto D (SD) and histidine-lactobionate (HL). The integrity of cold-stored rat and human islets of Langerhans has been examined using supravital stains and electron microscopy. In addition, the viability of cold-stored rat islets was tested by intraportal transplantation into syngeneic streptozotocin-induced diabetic recipients. The in vitro studies showed good preservation of islets stored in UW, SD, and HL for up to 72 hr. In comparison, storage for periods as short as 24 hr in HBSS markedly reduced islet integrity. The transplantation studies showed that rat islets cold stored in HBSS solution for 24 hr were not able to reverse experimental diabetes, whereas those stored in the other 3 solutions for 24 hr successfully reversed diabetes within 1 week of transplantation. After 48-hr cold storage, only islets preserved in HL solution were uniformly capable of producing functioning islet grafts. None of the tested solutions was able to maintain islet viability sufficiently to allow successful transplantation after more than 48 hr of cold storage. These experiments demonstrate that good islet viability is maintained for up to 24 hr of storage in UW, SD, and HL, and even after 48-hr cold storage in HL solution, whereas preservation in HBSS solution was deleterious to islet viability within 24 hr.

Key concepts: Viaspan, Islet, Cold storage, Transplantation, Balanced salt solution, Pancreatic islets, Streptozotocin, Andrology

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