Accelerating cryoprotectant diffusion kinetics improves cryopreservation of pancreatic islets

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

doi:10.1038/s41598-021-89853-6

Accelerating cryoprotectant diffusion kinetics improves cryopreservation of pancreatic islets

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

College of Medicine and Veterinary Medicine

Research output: Contribution to journal › Article › peer-review

Abstract / Description of output

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.

Original language	English
Pages (from-to)	10418
Journal	Scientific Reports
Volume	11
Issue number	1
DOIs	https://doi.org/10.1038/s41598-021-89853-6
Publication status	Published - 17 May 2021

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10.1038/s41598-021-89853-6

Dolezalova et al._ManuscriptTextAccepted author manuscript, 9.12 MB

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Dolezalova, N., Gruszczyk, A., Barkan, K., Gamble, J. A., Galvin, S., Moreth, T., O'Holleran, K., Mahbubani, K. T., Higgins, J. A., Gribble, F. M., Reimann, F., Surmacki, J., Andrews, S., Casey, J. J., Pampaloni, F., Murphy, M. P., Ladds, G., Slater, N. K. H., & Saeb-Parsy, K. (2021). Accelerating cryoprotectant diffusion kinetics improves cryopreservation of pancreatic islets. Scientific Reports, 11(1), 10418. https://doi.org/10.1038/s41598-021-89853-6

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title = "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 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.",

author = "Nikola Dolezalova and Anja Gruszczyk and Kerry Barkan and Gamble, {John A} and Sam Galvin and Till Moreth and Kevin O'Holleran and Mahbubani, {Krishnaa T} and Higgins, {Jackie A} and Gribble, {Fiona M} and Frank Reimann and Jakub Surmacki and Simon Andrews and Casey, {John J} and Francesco Pampaloni and Murphy, {Michael P} and Graham Ladds and Slater, {Nigel K H} and Kourosh Saeb-Parsy",

year = "2021",

month = may,

day = "17",

doi = "10.1038/s41598-021-89853-6",

language = "English",

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journal = "Scientific Reports",

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publisher = "Nature Publishing Group",

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Dolezalova, N, Gruszczyk, A, Barkan, K, Gamble, JA, Galvin, S, Moreth, T, O'Holleran, K, Mahbubani, KT, Higgins, JA, Gribble, FM, Reimann, F, Surmacki, J, Andrews, S, Casey, JJ, Pampaloni, F, Murphy, MP, Ladds, G, Slater, NKH & Saeb-Parsy, K 2021, 'Accelerating cryoprotectant diffusion kinetics improves cryopreservation of pancreatic islets', Scientific Reports, vol. 11, no. 1, pp. 10418. https://doi.org/10.1038/s41598-021-89853-6

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T1 - Accelerating cryoprotectant diffusion kinetics improves cryopreservation of pancreatic islets

AU - Dolezalova, Nikola

AU - Gruszczyk, Anja

AU - Barkan, Kerry

AU - Gamble, John A

AU - Galvin, Sam

AU - Moreth, Till

AU - O'Holleran, Kevin

AU - Mahbubani, Krishnaa T

AU - Higgins, Jackie A

AU - Gribble, Fiona M

AU - Reimann, Frank

AU - Surmacki, Jakub

AU - Andrews, Simon

AU - Casey, John J

AU - Pampaloni, Francesco

AU - Murphy, Michael P

AU - Ladds, Graham

AU - Slater, Nigel K H

AU - Saeb-Parsy, Kourosh

PY - 2021/5/17

Y1 - 2021/5/17

N2 - 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.

AB - 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.

U2 - 10.1038/s41598-021-89853-6

DO - 10.1038/s41598-021-89853-6

M3 - Article

C2 - 34001961

SN - 2045-2322

VL - 11

SP - 10418

JO - Scientific Reports

JF - Scientific Reports

IS - 1

ER -

Accelerating cryoprotectant diffusion kinetics improves cryopreservation of pancreatic islets

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