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A Simple High Efficiency Protocol for Pancreatic Islet Isolation from Mice
Published on: August 30, 2019
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Accelerating cryoprotectant diffusion kinetics improves cryopreservation of pancreatic islets
Nikola Dolezalova1,2, Anja Gruszczyk3, Kerry Barkan4,5
1Department of Surgery, University of Cambridge, Addenbrooke's Hospital, Hills Road, Box 202, Cambridge, CB2 0QQ, UK.
Scientific Reports
|May 18, 2021
Summary
Accelerating cryopreservation equilibration for pancreatic islets using higher temperatures improves survival. A new method combining dimethyl sulfoxide (DMSO) with trehalose enhances islet cryosurvival and function for diabetic treatments.
Area of Science:
- Regenerative Medicine
- Cell Biology
- Biotechnology
Background:
- Pancreatic islet transplantation is a potential treatment for diabetes.
- Current cryopreservation methods using dimethyl sulfoxide (DMSO) result in low islet survival rates.
- Improving islet cryopreservation is crucial for increasing their availability for therapeutic use.
Purpose of the Study:
- To accelerate the equilibration of small molecules into islets during cryopreservation.
- To develop a novel, improved cryopreservation protocol for pancreatic islets.
- To assess the viability and function of cryopreserved islets using the new protocol.
Main Methods:
- Mouse islets were equilibrated with small molecules at an accelerated rate by increasing incubation temperature to 37°C.
- A novel cryopreservation protocol was developed, combining DMSO with trehalose pre-incubation.
- Islet viability, ATP/ADP ratios, peptide secretion, cAMP response, and gene expression were analyzed post-cryopreservation.
Main Results:
- Accelerated equilibration from over 24 hours to 6 hours was achieved by increasing incubation temperature.
- The novel cryopreservation protocol demonstrated improved cryosurvival rates compared to standard methods.
- Cryopreserved islets exhibited enhanced ATP/ADP ratios, preserved peptide secretion and cAMP response, and a gene expression profile indicative of better cryoprotection.
Conclusions:
- Optimizing incubation temperature significantly accelerates islet equilibration for cryopreservation.
- The combined DMSO and trehalose cryopreservation method enhances islet viability and function.
- This improved protocol holds promise for increasing the supply of islets for transplantation and advancing cryopreservation techniques for other multicellular constructs.
