Studies of the Isolation and Viability of Human Islets of Langerhans1,2

Warnock, Garth L.; Ellis, Dwayne; Rajotte, Ray V.; Dawidson, Ingemar; Baekkeskov, Steinunn; Egebjerg, Jorn

doi:10.1097/00007890-198805000-00024

Cited by 63 publications

(30 citation statements)

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“…This observation is consistent with several previous reports (9,(15)(16)(17)19). In the present study, 7 hr of CIT was the time discriminating successful from failed isolations.…”

Section: Discussionsupporting

confidence: 94%

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Refinement of the Automated Method for Human Islet Isolation and Presentation of a Closed System for In Vitro Islet Culture

et al. 2004

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“…This observation is consistent with several previous reports (9,(15)(16)(17)19). In the present study, 7 hr of CIT was the time discriminating successful from failed isolations.…”

Section: Discussionsupporting

confidence: 94%

“…Of this volume only about 1/3 can be contained in the pancreas. The efficiency of collagenase present in the solution is almost negligible compared with that within the pancreas (7)(8)(9). A novel distension procedure was introduced together with techniques to detect and seal damaged areas of the pancreas.…”

mentioning

confidence: 99%

Refinement of the Automated Method for Human Islet Isolation and Presentation of a Closed System for In Vitro Islet Culture

et al. 2004

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“…Experimental Studies Islet preparations with low endocrine cell purity (<20% In the first study, the diabetic mice were divided into dithizone-stained cells) (33) were used in these studies.…”

Section: Methodsmentioning

confidence: 99%

Combination Therapy with Glucagon-Like Peptide-1 and Gastrin Induces β-Cell Neogenesis from Pancreatic Duct Cells in Human Islets Transplanted in Immunodeficient Diabetic Mice

Suarez-Pinzon

Lakey²,

Rabinovitch

2008

Cell Transplant

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Pancreatic islet transplantation as a treatment for type 1 diabetes is limited by human donor tissue availability. We investigated whether the β-cell mass in human isolated islets could be expanded by treatments with glucagon-like peptide-1 (GLP-1) and gastrin, peptides reported to stimulate β-cell growth in mice and rats with deficits in β-cell mass. Human islets with low endocrine cell purity (7% β-cells, 4% α-cells) and abundant exocrine cells (29% duct cells and 25% acinar cells) were implanted under the renal capsule of nonobese diabetic-severe combined immune deficiency (NOD-scid) mice made diabetic with streptozotocin. The mice were treated with GLP-1 and gastrin, separately and together, daily for 5 weeks. Blood glucose was significantly reduced only in mice implanted with human pancreatic cells and treated with GLP-1 plus gastrin. Correction of hyperglycemia was accompanied by increased insulin content in the human pancreatic cell grafts as well as by increased plasma levels of human C-peptide in the mice. Immunocytochemical examination revealed a fourfold increase in insulin-positive cells in the human pancreatic cell grafts in GLP-1 plus gastrin-treated mice, and most of this increase was accounted for by the appearance of cytokeratin 19-positive pancreatic duct cells expressing insulin. We conclude that combination therapy with GLP-1 and gastrin expands the β-cell mass in human islets implanted in immunodeficient diabetic mice, largely from pancreatic duct cells associated with the islets, and this is sufficient to ameliorate hyperglycemia in the mice.

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“…They were cold-preserved in University of Wisconsin solution or in Histidine ± Tryptophan ± Ketoglutarate (HTK) solution [15] for 3 to 20 h, the time between in situ flush and start of the isolation procedure at the central facility in Brussels. After collagenase digestion and Ficoll gradient centrifugation [16], the islet-enriched interphase was harvested, washed and cultured at 37°C under serum-free conditions for maximally 4 weeks [17]. If the preparations could not be used for transplantation within this period, they were cryopreserved according to the method of Rajotte et al [18].…”

Section: Methodsmentioning

confidence: 99%

Implantation of standardized beta-cell grafts in a liver segment of IDDM patients: graft and recipient characteristics in two cases of insulin-independence under maintenance immunosuppression for prior kidney graft

et al. 1998

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Islet transplantation can restore insulin production in insulin-dependent diabetic (IDDM) patients in whom this capacity had been lost for many years [1±7]. This observation has been made in kidney and liver recipients, where advantage is taken of the need for a continuous immune suppression. Survival of the grafts is variable, but generally less than 1 year. A prolonged beta-cell function with a state of insulin-independence after one year was achieved in 7 % of the cases recorded by the Islet Transplant Reg- Diabetologia (1998) Summary Islet allografts in insulin-dependent diabetic (IDDM) patients exhibit variable survival lengths and low rates of insulin-independence despite treatment with anti-T-cell antibodies and maintenance immunosuppression. Use of poorly characterized freshly isolated preparations makes it difficult to determine whether failures are caused by variations in donor tissue. This study assesses survival of standardized beta-cell allografts in C-peptide negative IDDM patients on maintenance immunosuppression following kidney transplantation and without receiving anti-T-cell antibodies or additional immunosuppression. Human islets were isolated from pancreatic segments after maximal 20 h cold-preservation. During culture, preparations were selected according to quality control tests and combined with grafts with standardized cell composition (³ 50 % beta cells), viability ( ³ 90 % ), total beta-cell number (1 to 2 × 10 6 /kg body weight) and insulin-producing capacity (2 to 4 nmol × graft ±1 × h ±1 ). Grafts were injected in a liver segment through the repermeabilized umbilical vein. After 2 weeks C-peptide positivity, four out of seven recipients became C-peptide negative; two of them were initially GAD 65 -antibody positive and exhibited a rise in titre during graft destruction. The other three patients remained C-peptide positive for more than 1 year, two of them becoming insulin-independent with near-normal fasting glycaemia and HbA 1 c ; they remained GAD 65 -and islet cell antibody negative. The three patients with surviving grafts presented a history of anti-thymocyte globulin therapy at kidney transplantation. Long-term surviving grafts increased C-peptide release following intravenous glucagon or oral glucose but not following intravenous glucose. Thus, cultured human beta-cells can survive for more than 1 year in IDDM patients on maintenance anti-rejection therapy for a prior kidney graft and without the need for an increased immunosuppression at the time of implantation. The use of functionally standardized beta-cell grafts helps to identify recipient and graft factors which influence their survival and metabolic effects. Insulin-independence can be achieved by injection of 1.5 million beta-cells per kg body weight in a liver segment. These beta-cell implants respond well to adenylcyclase activators but poorly to glucose. [Diabetologia (1998) 41: 452±459]

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Studies of the Isolation and Viability of Human Islets of Langerhans1,2

Cited by 63 publications

References 0 publications

Refinement of the Automated Method for Human Islet Isolation and Presentation of a Closed System for In Vitro Islet Culture

Refinement of the Automated Method for Human Islet Isolation and Presentation of a Closed System for In Vitro Islet Culture

Combination Therapy with Glucagon-Like Peptide-1 and Gastrin Induces β-Cell Neogenesis from Pancreatic Duct Cells in Human Islets Transplanted in Immunodeficient Diabetic Mice

Implantation of standardized beta-cell grafts in a liver segment of IDDM patients: graft and recipient characteristics in two cases of insulin-independence under maintenance immunosuppression for prior kidney graft

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