Isolation and culture of the islets of Langerhans of the guinea pig

Moskalewski, Stanisław

doi:10.1016/0016-6480(65)90059-6

Cited by 343 publications

(124 citation statements)

References 24 publications

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“…In principle, there are two ways of isolating pancreatic islets -the mechanical microdissection technique [10] and the enzymatic isolation procedure employing crude collagenase [ 12]. Although both techniques have been in use for several years, only a little is known about possible effects on the B-cells of the isolation procedure [8,13].…”

Section: Discussionmentioning

confidence: 99%

Function of microdissected pancreatic islets cultured in a chemically defined medium. I. Insulin content and release

et al. 1975

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Summary. Microdissected pancreatic islets from non-inbred ob/ ob-mice were cultured for 6 or 7 days in serum-free tissue culture medium 199. The insulin content of the islets decreased 60% during culture in 17 mM or 28 mM glucose and about 70% in the presence of 3.3 mM or 5.6 mM glucose. At the end of a culture period in high glucose, the sum of the insulin in the islet plus that in the culture medium was almost twice as high as the insulin content of fresh islets, indicating an active insulin biosynthesis. The maximal insulin response to glucose after culture in 17 mM or 28 mM glucose was about 40% of that in fresh islets; after culture in 3.3 mM glucose it was 10%. Half-maximal stimulation was observed at a glucose concentration of 5 mM for islets cultured with high glucose as compared to 9 mM for fresh islets. Like glucose, glibenclamide was a more effective insulin stimulator after culture with a high glucose concentration than with a low one. However, leucine-induced insulin release was not affected by the glucose concentration in the preceding culture medium. Whereas potentiation of glucose-stimulated release by arginine or dibutyryl-cAMP was independent of glucose concentration during the culture, theophylline released three times more insulin when the islets had been cultured with high glucose.

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Section: Discussionmentioning

confidence: 99%

Function of microdissected pancreatic islets cultured in a chemically defined medium. I. Insulin content and release

et al. 1975

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“…Islets were i. Lernmark: The Preparatlon of, and Studies on, Free Cell Suspensions isolated by eollagenase treatment of the pancreas [22,15]. In each experiment, the panereata from two animals were placed in KR-tIEPES medium containing 1 mg/ml BSA and 5 mM glucose.…”

Section: Animals and Isolation Of Isletsmentioning

confidence: 99%

“…Pancreatic islets isolated by freehand microdissection [7,11] or by collagenase treatment of the pancreas [22,15] are extensively used for studying the mechanisms of insulin and glucagon secretion, tIowever, the isolated islet is a complex model, not only because it contains at least four types of endocrine cells [14,10] but also because other structural elements are present, such as blood vessels, a surrounding connective tissue capsule and basement membranes. The latter structures constitute possible barriers which could well interfere with molecules' entering or leaving the incubated, isolated islet, especially as it has lost its capillary system.…”

mentioning

confidence: 99%

The preparation of, and studies on, free cell suspensions from mouse pancreatic islets

1974

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Summary. Pancreatic islets, isolated from the pancreas of obese-hyperglycemic mice, were used to prepare free islet cells in suspension. Batches of 100 islets were disrupted by meeha.nieal shaking for 10 see in a Ca~+-free HEPES-buffered Krebs-Ringer medium containing 1 mM EGTA. From 200--500 islets, about 2.2 • 106 cells could be obtained in suspension, corresponding to a yield of roughly 55% as calculated from the content of DNA in cells and islets. The isolated islet cells appeared wellpreserved in the light and the electron microscope and seemed to exhibit a high degree of viability as judged by viable cell counts, a radioactive assay for lysis and insulin release. Insulin release from islet cell suspensions, as calcuIated per cell, per content of DNA or insulin or per packed cell volume, was stimulated by glucose alone or iu combination with theophylline. The glucose response was low compared with that of intact isolated islets.

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“…Many of the available viability tests fail to exclude the possibility that the islet, as a microorgan with a subtle and complex organization, has been seriously injured. Surprisingly little is known about the effects of pancreas procurement, islet isolation, islet purifICation and islet banking procedures on the morphological integrity of the islet tissue 1,II,17,23,25,26,30,32,43,44. The relative paucity of studies dealing with the morphology of isolated islets probably reflects a complacent attitude that developed early after experience indicated that isolated rodent islet tissue was well preserved and that islet tissue appeared to be much more resistant to damage during the isolation procedure than acinar cells.…”

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confidence: 99%

Islet isolation assessment in man and large animals

et al. 1990

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The treatment of diabetes mellitus by transplantation of isolated pancreatic islets is an approach that remains the subject of research by a large number of investigators throughout the world. A crucial requirement for the success of this enterprise is the ability to prepare viable isolated islets in adequate quantity. Over the years numerous descriptions of procedures for islet isolation from the pancreas of experimental animals and of man have been advanced; each claiming to be an improvement on previous methods. Indeed, there certainly have been advances, although few techniques live up to the claims that are made in their support Part of the problem is the generally poor methodology

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Isolation and culture of the islets of Langerhans of the guinea pig

Cited by 343 publications

References 24 publications

Function of microdissected pancreatic islets cultured in a chemically defined medium. I. Insulin content and release

Function of microdissected pancreatic islets cultured in a chemically defined medium. I. Insulin content and release

The preparation of, and studies on, free cell suspensions from mouse pancreatic islets

Islet isolation assessment in man and large animals

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