Preparation of viable islet cells from dogs by a new method

Horaguchi, Atsushi; Merrell, Ronald C.

doi:10.2337/diabetes.30.5.455

Cited by 49 publications

(13 citation statements)

References 2 publications

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“…The method of Marchettiet al (1988) uses hand picking of porcine islets which gives islets of a lower purity than reported here and is laborious since upwards of 50,000 islets can be isolated from a single pig pancreas. With these modifications, our isolation procedure gives variable recoveries of insulin, glucagon and somatostatin, but the degree of islet purity, expressed as the insulin: amylase ratios, remains consistently high and much greater than the insulin: amylase ratios reported by other investigators for their islet isolation methods from large mammals (Horaguchi and Merrell 1981;Sutherland, Steffes, Bauer, McManus, Noe and Najarian 1974;Sutherland, Matas, Steffes and Najarian 1976). If porcine islets are to be considered for transplantation studies then it is important that the exocrine contamination is low (Gray, Sutton, McShane, Petersand Morris 1986).…”

Section: Discussionmentioning

confidence: 81%

Porcine Islet Isolation, Cellular Composition and Secretory Response

et al. 1989

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Porcine islets were isolated by infusion of a warm collagenase solution into whole pancreata followed by static incubation at 37 degrees C for 15 minutes. The pancreata were then chopped into small pieces and the free islets purified by filtration and centrifugation over a ficoll gradient. The insulin:amylase ratio of the islets compared to that in the intact pancreas was determined in 19 pancreata and indicates that the isolated islets were of a high degree of purity. The distribution of insulin, glucagon, somatostatin and pancreatic polypeptide containing cells in pig pancreas sections was compared with that in rat. Porcine islets were much smaller and less well defined than rat islets with infiltration of acinar material even into the islet core. The levels of insulin, glucagon and somatostatin in porcine pancreas and isolated porcine islets were measured using conventional radioimmunoassay techniques. The ratio of these hormones in the pancreas was 105.1:5.8:1 respectively, and in the islets 105.1:0.68:0.087 respectively. Fragmentation of the islets during the isolation may have led to the loss of glucagon and somatostatin-containing cells. Islets cultured overnight and tested with a range of glucose concentrations for one hour did not show a significant stimulation of insulin secretion in the presence of 8.3 mM or 16.7 mM glucose compared to that in 2.8 mM glucose. However freshly isolated islets challenged with 8.3 mM, 13.9 mM and 22.2 mM glucose showed a 1.8 fold, 2.0 fold and 2.3 fold response respectively, over that in 2.8 mM glucose.(ABSTRACT TRUNCATED AT 250 WORDS)

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Section: Discussionmentioning

confidence: 81%

Porcine Islet Isolation, Cellular Composition and Secretory Response

et al. 1989

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“…Canine endocrine cells were purified by ductal perfusion as previously described (10)(11)(12). Briefly, the pancreatic duct was perfused with 0.2% collagenase (type I; Sigma) for 25-40 min at 37°C.…”

Section: Methodsmentioning

confidence: 99%

Canine Islets in an Ultrafiltered Environment

Merrell¹,

Basadonna²

1985

Artificial Organs

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Molecular sieve membranes can protect pancreatic islets against immune recognition in diabetic patients treated by endocrine tissue replacement. These biocompatible membranes permit the passage of small peptides such as insulin, and preclude the diffusion of immunoglobulins and immunogenic molecules. However, the tissue must function indefinitely in an ultrafiltered environment determined by the sequestering membranes. The chronic perifusion of canine islet tissue was compared in ultrafiltered and microfiltered chambers. The biphasic pattern of insulin release by similar numbers of islets from the same pancrease preparation was not significantly different when tissue was cultured in a micro- or an ultrafiltered environment. The cumulative insulin output of the two systems was quite similar over 3 days of culture. Canine islet tissue can be sustained in an ultrafiltered environment with maintenance of insulin release to glucose stimulation, which is quantitatively similar to islet tissue maintained in chronic perifusion without ultrafiltration.

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“…Previously published reports in research dogs used trituration of pancreatic tissue by passage through metal needles and maceration of tissue with a blade to achieve mechanical disruption . Whereas these methods are less expensive and avoid the need for specialized equipment, they lead to excessive damage to islet cells and yield only limited numbers of nonsterile islets of variable purity that would not be sufficient for clinical use . Our laboratory has developed a modification of standard human methodology for islet harvest in dogs that can be performed with minimal specialized equipment .…”

Section: Donor Issuesmentioning

confidence: 99%

Pancreatic Islet Transplantation: From Dogs to Humans and Back Again

et al. 2014

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Pancreatic islet transplantation is a cell-based therapy that provides a potential cure for type 1 diabetes mellitus. After the introduction of an automated method for islet isolation and steroid-free immunosuppressive protocols, reversal of diabetes by islet transplantation is now performed at major human medical centers around the world. Despite extensive use of animal models in islet transplantation research, practical concerns have slowed the introduction of the technique into clinical veterinary practice and only a small number of studies have reported results of transplantation in dogs with spontaneously occurring diabetes mellitus; however, recent advances in islet isolation and encapsulation may make it possible to perform islet transplantation without immunosuppression in companion animals. This review summarizes experimental and clinical studies of pancreatic islet transplantation in dogs, including future directions for cell therapy in animals with naturally occurring disease.

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Preparation of viable islet cells from dogs by a new method

Cited by 49 publications

References 2 publications

Porcine Islet Isolation, Cellular Composition and Secretory Response

Porcine Islet Isolation, Cellular Composition and Secretory Response

Canine Islets in an Ultrafiltered Environment

Pancreatic Islet Transplantation: From Dogs to Humans and Back Again

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