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

Type 1 diabetes mellitus (T1DM) is one of the most frequently diagnosed endocrinopathies in dogs with an increasing prevalence, challenging life-long management and frequent complications. Pancreatic islet transplantation is a noninvasive and potentially curative treatment for T1DM. 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 (IEQ) 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 using 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 to 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.

Name thesis:degree_name

Master of Science

Level thesis:degree_level

masters

Discipline thesis:degree_discipline

Veterinary Clinical Sciences

Grantor dc:publisher

The Ohio State University

Year dc:date

2013

Author dc:creator

• Vrabelova, Daniela

Contributors dc:contributor

• Adin, Christopher

• Health Sciences
• Medicine
• pancreas
• diabetes
• dog
• isolation
• transplantation

Statement dc:rights

• unrestricted
• This thesis or dissertation is protected by copyright: all rights reserved. It may not be copied or redistributed beyond the terms of applicable copyright laws.

Language dc:language

English

Repository record dc:identifier

http://rave.ohiolink.edu/etdc/view?acc_num=osu1365099629

OAI identifier oai:identifier

oai:etd.ohiolink.edu:osu1365099629