Simplified Method to Isolate Highly Pure Canine Pancreatic Islets

Woolcott, Orison O.; Bergman, Richard N.; Richey, Joyce M.; Kirkman, Erlinda L.; Harrison, L. Nicole; Ionut, Viorica; Lottati, Maya; Zheng, Dan; Hsu, Isabel R.; Stefanovski, Darko; Kabir, Morvarid; Kim, Stella P.; Catalano, Karyn J.; Chiu, Jenny D.; Chow, Robert H.

doi:10.1097/mpa.0b013e318221fd0e

Cited by 16 publications

(23 citation statements)

References 66 publications

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“…Data on the size of pancreatic islets in healthy dogs are scarce. In a study on different isolation procedures of pancreatic islets in 14 healthy dogs the average dimension of islets varied from 50 to 325 µm, while in 75% of them it was 100 to 200 µm (17). In other studies on healthy beagles and cross-breed dogs the average dimension of pancreatic islets was 50-100 µm (8,10).…”

Section: Resultsmentioning

confidence: 97%

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Histopathological picture of pancreatic islets in dogs with congenital portosystemic shunt

Frymus¹,

Trębacz²,

Sobczak-Filipiak³

et al. 2020

Medycyna Weterynaryjna

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Hypoglycemia is frequently found in dogs suffering from portosystemic shunt (PSS). However, the mechanisms leading to abnormal blood glucose concentration in such dogs have not been studied. Therefore, investigations were undertaken to study the structure of pancreatic islets in 25 patients with congenital PSS (cPSS). Material for morphometry and histopathology was taken during the surgical closing of the abnormal blood vessel. A total of 75 islets (3 randomly chosen from each patient) were analyzed, and their average size was compared to reference values for dogs that were considered as 50-325 μm. The average size of 47 (63%) islets was below 50 μm, whereas the mean dimension of the largest islet did not exceeded 80 μm. The average islet size in all patients was 46 (SD 13.3) μm and in only 2 dogs (8%) all 3 analyzed islets were bigger than the lower reference value. Histopathological examination revealed cytoplasmic vesicles in pancreatic cells, as well as extracellular, homogenous, acidophilic deposits in pancreatic islets. These results indicate that in dogs suffering from cPSS the pancreatic islets are smaller than the reference values and their cells may contain abnormal structures.

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Section: Resultsmentioning

confidence: 97%

“…In young dogs (6 months), type B cells are most numerous, representing 52-82% of the total islet cells, and their amount is constant, in contrast to percentages of A and PP cells that differ depending on the region (9). The size of the islets in healthy dogs ranges from 50 to 325 µm (8,10,17).…”

Section: Praca Oryginalnamentioning

confidence: 99%

Histopathological picture of pancreatic islets in dogs with congenital portosystemic shunt

Frymus¹,

Trębacz²,

Sobczak-Filipiak³

et al. 2020

Medycyna Weterynaryjna

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“…Islets and adipose derived MSCs were Isolated and cultured from mice and dogs as previously reported . Purified human islets from nondiabetic subjects (Prodo Laboratories, Irvine, CA), and human adipose derived MSCs (P1, Lonza, Walkersville, MD) were identically cultured.…”

Section: Methodsmentioning

confidence: 99%

Durable Control of Autoimmune Diabetes in Mice Achieved by Intraperitoneal Transplantation of “Neo-Islets,” Three-Dimensional Aggregates of Allogeneic Islet and “Mesenchymal Stem Cells”

Westenfelder

Gooch²,

Hu³

et al. 2017

Stem Cells Translational Medicine

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Novel interventions that reestablish endogenous insulin secretion and thereby halt progressive end‐organ damage and prolong survival of patients with autoimmune Type 1 diabetes mellitus (T1DM) are urgently needed. While this is currently accomplished with allogeneic pancreas or islet transplants, their utility is significantly limited by both the scarcity of organ donors and life‐long need for often‐toxic antirejection drugs. Coadministering islets with bone marrow‐derived mesenchymal stem cells (MSCs) that exert robust immune‐modulating, anti‐inflammatory, anti‐apoptotic, and angiogenic actions, improves intrahepatic islet survival and function. Encapsulation of insulin‐producing cells to prevent immune destruction has shown both promise and failures. Recently, stem cell‐derived insulin secreting β‐like cells induced euglycemia in diabetic animals, although their clinical use would still require encapsulation or anti‐rejection drugs. Instead of focusing on further improvements in islet transplantation, we demonstrate here that the intraperitoneal administration of islet‐sized “Neo‐Islets” (NIs), generated by in vitro coaggregation of allogeneic, culture‐expanded islet cells with high numbers of immuno‐protective and cyto‐protective MSCs, resulted in their omental engraftment in immune‐competent, spontaneously diabetic nonobese diabetic (NOD) mice. This achieved long‐term glycemic control without immunosuppression and without hypoglycemia. In preparation for an Food and Drug Administration‐approved clinical trial in dogs with T1DM, we show that treatment of streptozotocin‐diabetic NOD/severe combined immunodeficiency mice with identically formed canine NIs produced durable euglycemia, exclusively mediated by dog‐specific insulin. We conclude that this novel technology has significant translational relevance for canine and potentially clinical T1DM as it effectively addresses both the organ donor scarcity (>80 therapeutic NI doses/donor pancreas can be generated) and completely eliminates the need for immunosuppression. Stem Cells Translational Medicine 2017;6:1631–1643

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“…The long digestion of the pancreas resulted in islets of high purity although the total number of free islets was low. Most isolated islets had a diameter smaller than 100 μm, which might be associated with islet fragmentation 11 . Considering these findings, enzyme blends of higher purity and more precise composition may be more appropriate to ensure consistent degradation activity without deteriorating islet integrity 3 .…”

Section: Discussionmentioning

confidence: 99%

Establishment of a protocol for the isolation of feline pancreatic islets

Brandão

Zini

Reusch

et al. 2018

Physiology & Behavior

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Diabetes mellitus is a common endocrinopathy in cats that is associated with pancreatic islets lesions. Research on isolated islets contributed to the understanding of the pathophysiology of human diabetes. Therefore, by improving the existing methods of isolation in cats, we aimed at increasing islet yield, purity and viability of feline isolated islets. Islet isolation was accomplished by pancreas perfusion with 80ml of Collagenase type IV through the pancreatic duct at the site of the major papilla. The enzymatic digestion was combined with mechanical disruption and controlled by dithizone staining. Purification was performed by filtration and handpicking. Purified islets were plated on extracellular matrix pre-coated plates and cultured for 48h. Feline islets with a high degree of viability and purity were isolated and cultured for the first time. Although the percentage of islet free from the acinar tissue relative to the total number of isolated islets was low compared to other species, the suggested protocol represents a promising progress in the procedure of islet isolation in cats.

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Simplified Method to Isolate Highly Pure Canine Pancreatic Islets

Cited by 16 publications

References 66 publications

Histopathological picture of pancreatic islets in dogs with congenital portosystemic shunt

Histopathological picture of pancreatic islets in dogs with congenital portosystemic shunt

Durable Control of Autoimmune Diabetes in Mice Achieved by Intraperitoneal Transplantation of “Neo-Islets,” Three-Dimensional Aggregates of Allogeneic Islet and “Mesenchymal Stem Cells”

Establishment of a protocol for the isolation of feline pancreatic islets

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