P-194

Long-term storage of pig pancreata in oxygen-charged perfluorohexyloctane improves islet survival after culture and islet in vitro function compared to perfluorodecalin

H. Brandhorst¹, W. Scott², M. Iken³, K. Papas², B. Theisinger⁴, O. Korsgren¹, D. Brandhorst^1
1 Uppsala University, Dept. of Immunology, Genetics & Pathology, Uppsala, Sweden; ² University of Minnesota, Schulze Diabetes Institute, Minneapolis, USA; ³ University Medical School, Dept. of Gastroenterology, Hepatology & Endocrinology, Hanover, Germany; ⁴ Novaliq GmbH, Heidelberg, Germany

Objective: As requested by the InternationalXenotransplantation Association the realization of islet xenotransplantationfor clinical trials requires breeding of suitable pigs in centralized biosecurefacilities to provide harvested pancreata to experienced isolation facilities.Pancreas transportation between donor center and islet production facility isfrequently associated with prolonged ischemia impairing islet isolation andtransplantation outcomes. The aim of the present study was to compare theefficiency of perfluorohexyloctan (F6H8), an oxygen carrier with lipophilicqualities and a low density, to perfluorodecalin (PFD) to prevent ischemicallyinduced damage during pancreas shipment prior to porcine islet isolation.

Methods: Retrieved pancreata weredissected into the connecting and splenic lobe, intraductally flushed withUW-solution and immersed alternately in either preoxygenated F6H8 or PFD.During preservation, the ratio of ATP:inorganic phosphate (Pi) content wascompared over time utilizing 31P-NMR spectroscopy. Pancreata were processedusing Serva NB1 collagenase and discontinuous Ficoll purification in a Cobe2991. Isolated islets were cultured for 3–4 days at 37°C and subjected toquality assessment.

Results: No effect was observed conceringdigestion variables, yield and purity of freshly isolated islets. Nevertheless,a significantly improved glucose-stimulated insulin response and increasedviability was detected in islets isolated from F6H8-preserved pancreata.Moreover, islet survival after culture was enhanced after storage in F6H8compared to PFD. NMR spectroscopy revealed that pancreatic lobes preserved inF6H8 exhibit consistently higher ATP:Pi ratios compared to PFD.

Conclusion: The present data suggest that F6H8 does not seem to increase yieldbut rather seem to improve quality of pig islets isolated after prolonged coldischemia.

Disclosure: B. Theisinger is employed by Novaliq GmbH, provider of F6H8

P-194

Long-term storage of pig pancreata in oxygen-charged perfluorohexyloctane improves islet survival after culture and islet in vitro function compared to perfluorodecalin

H. Brandhorst¹, W. Scott², M. Iken³, K. Papas², B. Theisinger⁴, O. Korsgren¹, D. Brandhorst^1
1 Uppsala University, Dept. of Immunology, Genetics & Pathology, Uppsala, Sweden; ² University of Minnesota, Schulze Diabetes Institute, Minneapolis, USA; ³ University Medical School, Dept. of Gastroenterology, Hepatology & Endocrinology, Hanover, Germany; ⁴ Novaliq GmbH, Heidelberg, Germany

Objective: As requested by the InternationalXenotransplantation Association the realization of islet xenotransplantationfor clinical trials requires breeding of suitable pigs in centralized biosecurefacilities to provide harvested pancreata to experienced isolation facilities.Pancreas transportation between donor center and islet production facility isfrequently associated with prolonged ischemia impairing islet isolation andtransplantation outcomes. The aim of the present study was to compare theefficiency of perfluorohexyloctan (F6H8), an oxygen carrier with lipophilicqualities and a low density, to perfluorodecalin (PFD) to prevent ischemicallyinduced damage during pancreas shipment prior to porcine islet isolation.

Methods: Retrieved pancreata weredissected into the connecting and splenic lobe, intraductally flushed withUW-solution and immersed alternately in either preoxygenated F6H8 or PFD.During preservation, the ratio of ATP:inorganic phosphate (Pi) content wascompared over time utilizing 31P-NMR spectroscopy. Pancreata were processedusing Serva NB1 collagenase and discontinuous Ficoll purification in a Cobe2991. Isolated islets were cultured for 3–4 days at 37°C and subjected toquality assessment.

Results: No effect was observed conceringdigestion variables, yield and purity of freshly isolated islets. Nevertheless,a significantly improved glucose-stimulated insulin response and increasedviability was detected in islets isolated from F6H8-preserved pancreata.Moreover, islet survival after culture was enhanced after storage in F6H8compared to PFD. NMR spectroscopy revealed that pancreatic lobes preserved inF6H8 exhibit consistently higher ATP:Pi ratios compared to PFD.

Conclusion: The present data suggest that F6H8 does not seem to increase yieldbut rather seem to improve quality of pig islets isolated after prolonged coldischemia.

Disclosure: B. Theisinger is employed by Novaliq GmbH, provider of F6H8