The scarcity of available tissue for transplantation in diabetes and the need for multiple donors make it mandatory to use an optimal cryopreservation method that allows maximal recovery and preservation of β-cell function. We have developed a method to cryopreserve islets with excellent survival of endocrine cells. Current methods use DMSO as cryoprotectant. Our method involves introducing both DMSO and the disaccharide trehalose into the cells during cooling. Uptake and release of trehalose occurred during the thermotropic lipid-phase transition measured in pancreatic endocrine cells between 5° and 9°C, using [14C]trehalose. Recovery of adult islets after cryopreservation with 300 mmol/l trehalose was 92 vs. 58% using DMSO alone. In vitro function, in terms of insulin content and release in response to secretagogues, was indistinguishable from fresh islets. Grafts from islets cryopreserved with trehalose contained 14-fold more insulin than grafts from islets cryopreserved without trehalose. Results with human fetal islet-like cell clusters (ICCs) were more pronounced: recovery from cryopreservation was 94%, compared with 42% without trehalose. Complete functionality of fetal cells was also restored; tritiated thymidine incorporation and insulin content and release were similar to fresh tissue. After transplantation in nude mice, there was a 15-fold increase in insulin content of grafts from ICCs cryopreserved with trehalose compared with ICCs cryopreserved without trehalose. Thus, the addition of trehalose to cryopreservation protocols leads to previously unobtainable survival rates of human pancreatic endocrine tissue.
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March 01 1997
Trehalose: A Cryoprotectant That Enhances Recovery and Preserves Function of Human Pancreatic Islets After Long-Term Storage
Gillian M Beattie;
Gillian M Beattie
Whittier Institute, Department of Pediatrics, University of California at San Diego School of Medicine
La Jolla
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John H Crowe;
John H Crowe
Section of Molecular and Cellular Biology, University of California at Davis
Davis, California
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Ana D Lopez;
Ana D Lopez
Whittier Institute, Department of Pediatrics, University of California at San Diego School of Medicine
La Jolla
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Vincenzo Cirulli;
Vincenzo Cirulli
Whittier Institute, Department of Pediatrics, University of California at San Diego School of Medicine
La Jolla
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Camillo Ricordi;
Camillo Ricordi
Diabetes Research Institute, University of Miami
Miami, Florida
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Alberto Hayek
Alberto Hayek
Whittier Institute, Department of Pediatrics, University of California at San Diego School of Medicine
La Jolla
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Address correspondence and reprint requests to Dr. A. Hayek, Whittier Institute, Department of Pediatrics, 9894 Genesee Ave., La Jolla, CA 92037. [email protected].
1
ANOVA, analysis of variance; FTIR, Fourier transform infrared; ICC, islet-like cell clusters; NIH, National Institutes of Health; RIA, radioimmunoassay.
Diabetes 1997;46(3):519–523
Article history
Received:
October 18 1996
Revision Received:
December 03 1996
Accepted:
December 03 1996
PubMed:
9032112
Citation
Gillian M Beattie, John H Crowe, Ana D Lopez, Vincenzo Cirulli, Camillo Ricordi, Alberto Hayek; Trehalose: A Cryoprotectant That Enhances Recovery and Preserves Function of Human Pancreatic Islets After Long-Term Storage. Diabetes 1 March 1997; 46 (3): 519–523. https://doi.org/10.2337/diab.46.3.519
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