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Domest Anim Endocrinol. 2014 Apr;47:119-26. doi: 10.1016/j.domaniend.2013.01.006. Epub 2013 Feb 15.

Evaluation of a high-yield technique for pancreatic islet isolation from deceased canine donors.

Author information

1
Department of Veterinary Clinical Sciences and Surgery, The Ohio State University, Columbus, OH 43210, USA.
2
Department of Veterinary Clinical Sciences and Surgery, The Ohio State University, Columbus, OH 43210, USA. Electronic address: adin.1@osu.edu.
3
Department of Surgery, The Ohio State University, Columbus, OH 43210, USA.

Abstract

Type 1 diabetes mellitus is one of the most frequently diagnosed endocrinopathies in dogs, and prevalence continues to increase. Pancreatic islet transplantation is a noninvasive and potentially curative treatment for type 1 diabetes mellitus. Institution of this treatment in dogs will require a readily available source of canine islets. We hypothesized that clinically acceptable islet yield and purity could be achieved by using deceased canine donors and standard centrifugation equipment. Pancreata were procured from dogs euthanized for reasons unrelated to this study. Initial anatomic studies were performed to evaluate efficacy of pancreatic perfusion. Infusion into the accessory pancreatic duct resulted in perfusion of approximately 75% of the pancreas. Additional cannulation of the distal right limb of the pancreas allowed complete perfusion. Collagenase digestion was performed with a Ricordi chamber and temperature-controlled perfusion circuit. Islets were separated from the exocrine tissue with the use of a discontinuous density gradient and a standard laboratory centrifuge. After isolation, islet yield was calculated and viability was assessed with dual fluorescent staining techniques. Islet isolation was completed in 6 dogs. Median (interquartile range) islet yield was 36,756 (28,527) islet equivalents per pancreas. A high degree of islet purity (percentage of endocrine tissue; 87.5% [10%]) and viability (87.4% [12.4%]) were achieved. The islet yield achieved with this technique would require approximately 1 pancreas per 5 kg body weight of the recipient dog. Purity and viability of the isolated islets were comparable with those achieved in human islet transplantation program. According to initial results, clinically relevant islet yield and quality can be obtained from deceased canine donors with the use of standard laboratory equipment.

KEYWORDS:

Canine islets; Digestion; Pancreas; Ricordi chamber

PMID:
23428563
DOI:
10.1016/j.domaniend.2013.01.006
[Indexed for MEDLINE]
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