Author:
Dolezalova Nikola,Gruszczyk Anja,Barkan Kerry,Gamble John A.,Galvin Sam,Moreth Till,O’Holleran Kevin,Mahbubani Krishnaa T.,Higgins Jackie A.,Gribble Fiona M.,Reimann Frank,Surmacki Jakub,Andrews Simon,Casey John J.,Pampaloni Francesco,Murphy Michael P.,Ladds Graham,Slater Nigel K. H.,Saeb-Parsy Kourosh
Abstract
AbstractCryopreservation 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.
Funder
W. D. Armstrong Fund
MRC UK
Wellcome Trust Investigator award
Leverhulme Trust
BBSRC iCASE PhD studentship partnered with AstraZeneca, Wellcome Trust
Medical Research Council (MRC) Metabolic Diseases Unit
MRC “Enhancing UK clinical research” grant
EU Horizon2020 project LSFM4LIFE
Publisher
Springer Science and Business Media LLC
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