Accelerating cryoprotectant diffusion kinetics improves cryopreservation of pancreatic islets
Abstract Cryopreservation offers the potential to increase the availability of pancreatic islets for treatment of diabetic patients. However, current protocols, which use dimethyl sulfoxide (DMSO), lead to poor cryosurvival of islets. We demonstrate that equilibration of mouse islets with small mole...
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doaj-b718aa2f9b114fbdad8edf4f86a67ef92021-05-23T11:33:13ZengNature Publishing GroupScientific Reports2045-23222021-05-0111111810.1038/s41598-021-89853-6Accelerating cryoprotectant diffusion kinetics improves cryopreservation of pancreatic isletsNikola Dolezalova0Anja Gruszczyk1Kerry Barkan2John A. Gamble3Sam Galvin4Till Moreth5Kevin O’Holleran6Krishnaa T. Mahbubani7Jackie A. Higgins8Fiona M. Gribble9Frank Reimann10Jakub Surmacki11Simon Andrews12John J. Casey13Francesco Pampaloni14Michael P. Murphy15Graham Ladds16Nigel K. H. Slater17Kourosh Saeb-Parsy18Department of Surgery, University of Cambridge, Addenbrooke’s HospitalMRC Mitochondrial Biology Unit, University of Cambridge, Keith Peters BuildingDepartment of Pharmacology, University of CambridgeDepartment of Surgery, University of Cambridge, Addenbrooke’s HospitalInstitute of Metabolic Science, University of Cambridge, Addenbrooke’s HospitalBuchman Institute for Molecular Life Science, Goethe University FrankfurtCambridge Advanced Imaging Centre, University of CambridgeDepartment of Surgery, University of Cambridge, Addenbrooke’s HospitalDepartment of Surgery, University of Cambridge, Addenbrooke’s HospitalInstitute of Metabolic Science, University of Cambridge, Addenbrooke’s HospitalInstitute of Metabolic Science, University of Cambridge, Addenbrooke’s HospitalCavendish Laboratory, University of CambridgeBioinformatics Group, The Babraham InstituteDepartment of Clinical Surgery, University of Edinburgh, Royal Infirmary of EdinburghBuchman Institute for Molecular Life Science, Goethe University FrankfurtMRC Mitochondrial Biology Unit, University of Cambridge, Keith Peters BuildingDepartment of Pharmacology, University of CambridgeDepartment of Chemical Engineering and Biotechnology, University of CambridgeDepartment of Surgery, University of Cambridge, Addenbrooke’s HospitalAbstract Cryopreservation offers the potential to increase the availability of pancreatic islets for treatment of diabetic patients. However, current protocols, which use dimethyl sulfoxide (DMSO), lead to poor cryosurvival of islets. We demonstrate that equilibration of mouse islets with small molecules in aqueous solutions can be accelerated from > 24 to 6 h by increasing incubation temperature to 37 °C. We utilize this finding to demonstrate that current viability staining protocols are inaccurate and to develop a novel cryopreservation method combining DMSO with trehalose pre-incubation to achieve improved cryosurvival. This protocol resulted in improved ATP/ADP ratios and peptide secretion from β-cells, preserved cAMP response, and a gene expression profile consistent with improved cryoprotection. Our findings have potential to increase the availability of islets for transplantation and to inform the design of cryopreservation protocols for other multicellular aggregates, including organoids and bioengineered tissues.https://doi.org/10.1038/s41598-021-89853-6 |
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DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Nikola Dolezalova Anja Gruszczyk Kerry Barkan John A. Gamble Sam Galvin Till Moreth Kevin O’Holleran Krishnaa T. Mahbubani Jackie A. Higgins Fiona M. Gribble Frank Reimann Jakub Surmacki Simon Andrews John J. Casey Francesco Pampaloni Michael P. Murphy Graham Ladds Nigel K. H. Slater Kourosh Saeb-Parsy |
spellingShingle |
Nikola Dolezalova Anja Gruszczyk Kerry Barkan John A. Gamble Sam Galvin Till Moreth Kevin O’Holleran Krishnaa T. Mahbubani Jackie A. Higgins Fiona M. Gribble Frank Reimann Jakub Surmacki Simon Andrews John J. Casey Francesco Pampaloni Michael P. Murphy Graham Ladds Nigel K. H. Slater Kourosh Saeb-Parsy Accelerating cryoprotectant diffusion kinetics improves cryopreservation of pancreatic islets Scientific Reports |
author_facet |
Nikola Dolezalova Anja Gruszczyk Kerry Barkan John A. Gamble Sam Galvin Till Moreth Kevin O’Holleran Krishnaa T. Mahbubani Jackie A. Higgins Fiona M. Gribble Frank Reimann Jakub Surmacki Simon Andrews John J. Casey Francesco Pampaloni Michael P. Murphy Graham Ladds Nigel K. H. Slater Kourosh Saeb-Parsy |
author_sort |
Nikola Dolezalova |
title |
Accelerating cryoprotectant diffusion kinetics improves cryopreservation of pancreatic islets |
title_short |
Accelerating cryoprotectant diffusion kinetics improves cryopreservation of pancreatic islets |
title_full |
Accelerating cryoprotectant diffusion kinetics improves cryopreservation of pancreatic islets |
title_fullStr |
Accelerating cryoprotectant diffusion kinetics improves cryopreservation of pancreatic islets |
title_full_unstemmed |
Accelerating cryoprotectant diffusion kinetics improves cryopreservation of pancreatic islets |
title_sort |
accelerating cryoprotectant diffusion kinetics improves cryopreservation of pancreatic islets |
publisher |
Nature Publishing Group |
series |
Scientific Reports |
issn |
2045-2322 |
publishDate |
2021-05-01 |
description |
Abstract Cryopreservation offers the potential to increase the availability of pancreatic islets for treatment of diabetic patients. However, current protocols, which use dimethyl sulfoxide (DMSO), lead to poor cryosurvival of islets. We demonstrate that equilibration of mouse islets with small molecules in aqueous solutions can be accelerated from > 24 to 6 h by increasing incubation temperature to 37 °C. We utilize this finding to demonstrate that current viability staining protocols are inaccurate and to develop a novel cryopreservation method combining DMSO with trehalose pre-incubation to achieve improved cryosurvival. This protocol resulted in improved ATP/ADP ratios and peptide secretion from β-cells, preserved cAMP response, and a gene expression profile consistent with improved cryoprotection. Our findings have potential to increase the availability of islets for transplantation and to inform the design of cryopreservation protocols for other multicellular aggregates, including organoids and bioengineered tissues. |
url |
https://doi.org/10.1038/s41598-021-89853-6 |
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