Insulin and Glucagon Secretion from Rat Islets Maintained in a Tissue Culture-Perifusion System

Gingerich, Ronald L.; Aronoff, Stephen L.; Kipnis, David; Lacy, Paul E.

doi:10.2337/diab.28.4.276

Cited by 12 publications

(3 citation statements)

References 4 publications

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“…In vitro function of freshly isolated and cultured islets was assessed by applying glucose perifusions as previously described (Gingerich et al, 1979). Aliquots of 100 IEQ were loaded into perifusion chambers and subjected to a dynamic glucose challenge after overnight culture at 37°C and an additional equilibration period of 2 h in basal glucose perifusion medium (2.7 mM).…”

Section: Islet In Vitro Functionmentioning

confidence: 99%

Islet Isolation from the Pancreas of Large Mammals and Humans: 10 Years of Experience

Brandhorst¹,

Brandhorst²,

Hering³

et al. 2009

Exp Clin Endocrinol Diabetes

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Despite experience, that has been obtained in the field of islet isolation in large mammals for about the past 20 years, allo- and autotransplantation of both human and porcine islets are still not routine procedures. The reasons for islet isolation and purification associated problems, which prevent a continuous isolation success and regular islet transplantation, can be categorized to variables related to 1) pancreas donor; 2) pancreas procurement; 3) isolation-techniques and 4) purification techniques. The development of porcine and human islet isolation was carried out by the authors since 1986. During this period several techniques for the isolation and purification of human and porcine islets were compared with regard to their influence on islet isolation outcome, integrity of islet morphology, islet in-vitro and in-vivo function: sequential vs continuous digestion-filtration, counterflow elutriation vs neutral density separation, conventional density gradient centrifugation vs Cobe cell separation and discontinuous vs continuous gradient purification. Furthermore, we analysed the influence of donor factors and variables related to organ procurement on islet isolation success. From this experience we concluded that successful porcine islet isolation is possible if islets of adult donors are isolated utilizing the continuous digestion-filtration following distension with UW-solution prior to a neutral density gradient purification on a Cobe. Human islets can be successfully isolated by digestion-filtration and purified utilizing a continuous Ficoll-sodium-diatrizoate gradient on a Cobe when intact pancreata are harvested after a secondary pancreatic warm ischemia time < 20 min from adult donors > 30 years.

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Section: Islet In Vitro Functionmentioning

confidence: 99%

Islet Isolation from the Pancreas of Large Mammals and Humans: 10 Years of Experience

Brandhorst¹,

Brandhorst²,

Hering³

et al. 2009

Exp Clin Endocrinol Diabetes

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“…Islet insulin secretion was studied using a multi-channel perifusion system, modified from Gingerich, Aronoff, Kipnis and Lacy (1979). Carefully counted numbers of islets from each size group were perifused at 37° C in 2 ml plastic syringe chambers containing a supporting matrix of 2.5 mg Cytodex-1 microcarrier beads, through which medium was pumped at 1.0 ml/min.…”

Section: Insulin Secretion From Isletsmentioning

confidence: 99%

Standardization of Insulin Secretion from Pancreatic Islets: Validation of a DNA Assay

Hopcroft¹,

Mason²,

Scott³

1985

Horm Metab Res

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The use of islet DNA content to standardize insulin secretion rates from pancreatic islets of different sizes has been studied. Isolated intact islets were sorted into 4 size categories and perifused with 22 mM glucose, collecting effluent in 5 min fractions for insulin RIA. DNA content of perifused islets was measured by fluorometric assay, and insulin secretion expressed as pmoles/ug DNA/unit time. For islets with diameters less than 300 u (1) insulin secretion was proportional to islet size; (2) insulin release per islet and islet DNA content were strongly correlated; (3) when expressed as a function of DNA content, insulin secretion from different sized islets was not significantly different. These relationships did not continue for very large islets (above 300 u) suggesting a limiting islet size for insulin secretion in vitro. The data demonstrates that expression of insulin secretion from pancreatic islets with diameters less than 300 u, as a function of their DNA content standardizes secretion irrespective of islet size and number, and should allow direct comparison of secretory responses between different islet tissue preparations.

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“…Recent technological innovations and the advent of microfluidics have brought dynamic perifusion to the forefront of islet and SC-β functional assessment. These systems allow for high-throughput study of basic function and more complex investigations into the dynamic effects of drugs and secretagogues on islet multihormonal secretion patterns 16–25. Despite these advances, the systems are prone to technical and budgetary limitations.…”

Section: Introductionmentioning

confidence: 99%

A static glucose-stimulated insulin secretion (sGSIS) assay that is significantly predictive of time to diabetes reversal in the human islet bioassay

Molano,

Pileggi,

Tse

et al. 2024

BMJ Open Diab Res Care

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IntroductionStatic incubation (static glucose-stimulated insulin secretion, sGSIS) is a measure of islet secretory function. The Stimulation Index (SI; insulin produced in high glucose/insulin produced in low glucose) is currently used as a product release criterion of islet transplant potency.Research design and methodsOur hypothesis was that the Delta, insulin secreted in high glucose minus insulin secreted in low glucose, would be more predictive. To evaluate this hypothesis, sGSIS was performed on 32 consecutive human islet preparations, immobilizing the islets in a slurry of Sepharose beads to minimize mechanical perturbation. Simultaneous full-mass subrenal capsular transplants were performed in chemically induced diabetic immunodeficient mice. Logistic regression analysis was used to determine optimal cut-points for diabetes reversal time and the Fisher Exact Test was used to assess the ability of the Delta and the SI to accurately classify transplant outcomes. Receiver operating characteristic curve analysis was performed on cut-point grouped data, assessing the predictive power and optimal cut-point for each sGSIS potency metric. Finally, standard Kaplan-Meier-type survival analysis was conducted.ResultsIn the case of the sGSIS the Delta provided a superior islet potency metric relative to the SI.ConclusionsThe sGSIS Delta value is predicitive of time to diabetes reversal in the full mass human islet transplant bioassay.

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Insulin and Glucagon Secretion from Rat Islets Maintained in a Tissue Culture-Perifusion System

Cited by 12 publications

References 4 publications

Islet Isolation from the Pancreas of Large Mammals and Humans: 10 Years of Experience

Islet Isolation from the Pancreas of Large Mammals and Humans: 10 Years of Experience

Standardization of Insulin Secretion from Pancreatic Islets: Validation of a DNA Assay

A static glucose-stimulated insulin secretion (sGSIS) assay that is significantly predictive of time to diabetes reversal in the human islet bioassay

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