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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Autores principales:	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
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/b718aa2f9b114fbdad8edf4f86a67ef9
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spelling	oai:doaj.org-article:b718aa2f9b114fbdad8edf4f86a67ef92021-12-02T14:58:53ZAccelerating cryoprotectant diffusion kinetics improves cryopreservation of pancreatic islets10.1038/s41598-021-89853-62045-2322https://doaj.org/article/b718aa2f9b114fbdad8edf4f86a67ef92021-05-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-89853-6https://doaj.org/toc/2045-2322Abstract 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.Nikola DolezalovaAnja GruszczykKerry BarkanJohn A. GambleSam GalvinTill MorethKevin O’HolleranKrishnaa T. MahbubaniJackie A. HigginsFiona M. GribbleFrank ReimannJakub SurmackiSimon AndrewsJohn J. CaseyFrancesco PampaloniMichael P. MurphyGraham LaddsNigel K. H. SlaterKourosh Saeb-ParsyNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-18 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q 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
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.
format	article
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
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 Portfolio
publishDate	2021
url	https://doaj.org/article/b718aa2f9b114fbdad8edf4f86a67ef9
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Accelerating cryoprotectant diffusion kinetics improves cryopreservation of pancreatic islets

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