Insulin Release, Insulin Content, and Glucose Utilization in Isolated Mouse Islets Maintained in Tissue Culture

Eff, C.

doi:10.1530/acta.0.0870535

Cited by 8 publications

(2 citation statements)

References 0 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…28 -3°- 32 Lacy and Gingerich 33 showed that the acute response to a glucose stimulus (5 g/L) was better preserved in isolated adult islets that had been cultured in 2.5 g/L glucose than in islets maintained in 1 g/L glucose. Eff 34 showed that glucose-stimulated insulin release was higher for cultures of isolated adult mouse islets that had been grown for 6 days at 27.8 mM glucose and then 4 days at 6.1 mM glucose than islets maintained at 6.1 mM glucose for the entire 10 days. Hellerstrom et al 35 showed that islets obtained with mild collagenase digestion of fetal rat pancreata and cultured for 8 days in medium containing 11.1 or 22.2 mM glucose responded to an acute glucose stimulus (16.7 mM) whereas there was no response by islets cultured in 5.5 or 2.8 mM glucose.…”

Section: Discussionmentioning

confidence: 99%

Organ Culture of Fetal Mouse Pancreas: The Effect of Culture Conditions on Insulin and Glucagon Secretion

et al. 1981

View full text Add to dashboard Cite

Seventeen-day-old mouse pancreas was grown in organ culture under various culture conditions and the insulin released into the medium, either chronically or during acute incubations, was measured. Three media, DME, RPMI 1640, and TC199, were tested, each at glucose concentrations of 1, 2, 3, and 4 g/L. Fetal pancreata maintained in RPM11640 at 4 g/L glucose secreted the most insulin over a 25-day period. Tissue grown in RPMI 1640 and DME secreted more insulin in higher glucose concentrations, but explants grown in TC199 secreted low levels of insulin regardless of glucose concentration. Fetal pancreata grown in medium supplemented with 5% FCS secreted less insulin in acute incubations than tissues grown in 10%, 15%, 20%, or 30% FCS, which secreted equivalent amounts. There was no difference in the amount of insulin released by islets grown in media supplemented with 15% FCS from four different batches.Although fetal islets grown in high glucose concentrations generally secreted more insulin (and less glucagon) than "normoglycemic" controls, their ability to release insulin in subsequent acute incubations was impaired. Explants, which had been maintained for 2 wk in 2.5 or 4 g/L glucose and a further 3 days in normoglycemic medium, secreted less insulin and glucagon in acute release exeriments than tissue maintained in 1 g/L glucose throughout.Light microscopy of cultured fetal pancreas showed that tissue remained viable at all glucose concentrations tested and differences in response could not be attributed to selective necrosis. Tranplantation of cultured islets indicated that tissue grown under normoglycemic conditions functioned better than tissue grown in hyperglycemic media. DIABETES 30:804-812, October 1981.

show abstract

Section: Discussionmentioning

confidence: 99%

Organ Culture of Fetal Mouse Pancreas: The Effect of Culture Conditions on Insulin and Glucagon Secretion

et al. 1981

View full text Add to dashboard Cite

show abstract

“…While direct injection worked well for mouse islet isolation, 40 it had not been effective in our previous attempts in the dog, human, or monkey pancreas and was not further evaluated. Ductal injection had the problems previously described plus the obvious drawback of low islet yields.…”

mentioning

confidence: 99%

Isolating the Elusive Islet

et al. 1980

View full text Add to dashboard Cite

The lack of a technique that allows mass isolation of intact, viable human islets is part of the reason that islet transplantation has not become available to the human diabetic. This report outlines the history of islet isolation and presents two new technical modifications that have been developed in the dog. Many of the current problems in islet isolation are presented, including the difficulty in obtaining enough human pancreatic tissue with minimal warm-ischemia time; inadequate distention of the pancreas to provide sufficient disruption for maximal enzymatic reaction to release intact islets; inefficient chopping methods; the use of collagenase of variable composition; different digestion methods for obtaining isolated islets; and inefficient methods for separating and purifying the islets from the ductal, acinar, and fibrous components. The first new modification involves distention of the dog pancreas through the venous system of the gland rather than the ductal system. This results in improved intralobular disruption, which improved the yield of isolated dog islets by permitting more efficient collagenase digestion. The second new modification eliminates the concept of isolating intact islets: the dog pancreas is digested by trypsin to a single-cell preparation that is partially purified by Ficoll gradients; further purification of the endocrine cells results from selective aggregation using rotational culture. This process produces pseudoislets that contain all the islet cell types and can be kept in culture for up to 4 wk, releasing their hormones in response to appropriate stimuli. These modifications may assist in the struggle to isolate the elusive human islet for safe and effective islet transplantation in the diabetic patient.

show abstract

Organ culture of the islets of Langerhans from young and senescent rats

et al. 1980

View full text Add to dashboard Cite

The B-cells of the endocrine pancreas constitute an adequate model for in vitro study of the aging process in highly differentiated cells. In the present study, collagenase-isolated islets of Langerhans from young and senescent rats were cultured up to 28 days. The response of the B-cells to the stimulatory conditions of the culture medium involved the nucleus, ribosomes, endoplasmic reticulum, Golgi apparatus, and secretory granules. Correlated data from light microscopy, electron microscopy, and insulin radioimmunoassay show that the differentiation and function of senescent B-cells are maintained in culture, as it has been proven for the B-cells of younger animals. On the other hand, signs of cytological deficiency not directly concerned with the specific function of B-cells were observed: abnormal mitochondria and lysosomes are more numerous in the senescent B-cells. The proliferative capacity of the B-cells of aged rats is reduced.

show abstract

Insulin Release, Insulin Content, and Glucose Utilization in Isolated Mouse Islets Maintained in Tissue Culture

Cited by 8 publications

References 0 publications

Organ Culture of Fetal Mouse Pancreas: The Effect of Culture Conditions on Insulin and Glucagon Secretion

Organ Culture of Fetal Mouse Pancreas: The Effect of Culture Conditions on Insulin and Glucagon Secretion

Isolating the Elusive Islet

Organ culture of the islets of Langerhans from young and senescent rats

Contact Info

Product

Resources

About