Summary
Microdissected pancreatic islets from non-inbredob/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.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
Kostianovsky, M., Lacy, P.E., Greider, M.H., Still, M.F.: Long term (15 days) incubation of islets of Langerhans isolated from adult rats and mice. Lab. Invest.27, 53–61 (1972)
Andersson, A.: Long-term effects of glucose on insulin release and glucose oxidation by mouse pancreatic islets maintained in tissue culture. Biochem. J.140, 377–382 (1974)
Ziegler, B., Hahn, H.-J., Fiedler, H.: Das kultivierte Pankreas als ein Modell für Untersuchungen zur Differenzierung und Hormonsekretion. Biol. Zbl.93, 467–479 (1974)
Andersson, A., Hellerström, C.: Metabolic characteristics of isolated pancreatic islets in tissue culture. Diabetes21 (Suppl. 2), 546–554 (1972)
Ziegler, B., Ziegler, M., Hahn, H.-J., Fiedler, H.: Untersuchungen an Langerhans'schen Inseln in vitro. X. Kultivierung von Langerhans'schen Inseln der Wistarratte und deren Insulinsekretionsverhalten. Acta biol. med. germ.32, 503–511 (1974)
Andersson, A.: Effects of glucose on the structure and metabolism of isolated pancreatic islets maintained in tissue culture. Thesis, Uppsala (1973)
Salk, J., Youngner, J.S., Ward, E.: Method of preparing mixture 199. Amer. J. Hyg.60, 225–230 (1954)
Lernmark, Å.: Isolated mouse islets as a model for studying insulin release. Acta diabet. lat.8, 649–679 (1971)
Hellman, B.: Studies in obese-hyperglycemic mice. Ann. N. Y. Acad. Sci.131, 541–558 (1965)
Hellerström, C.: A method for the microdissection of intact pancreatic islets of mammals. Acta endocr. (Kbh.)45, 122–132 (1964)
Hales, C.N., Randle, P.J.: Immunoassay of insulin with insulinantibody precipitate. Biochem. J.88, 137–146 (1963)
Moskalewski, S.: Isolation and culture of the islets of Langerhans of the guinea pig. Gen. comp. Endocr.5, 342–353 (1965)
Hellman, B., Lernmark, Å.: Methods for the isolation of cells with special reference to the pancreatic islets. In: Recent advances in quantitative histo- and cytochemistry, (eds. U.C. Dubach, U. Schmidt), pp. 91–107. Bern: Hans Huber Publishers 1971
Boder, G.B., Root, M.A., Chance, R.E., Johnson, I.S.: Extended production of insulin by isolated rabbit pancreatic islets; Evidence for biosynthesis of insulin. Proc. Soc. exp. Biol. (N. Y.)131, 507–513 (1969)
Krause, U., Puchinger, H., Wacker, A.: Inhibition of glucose-induced insulin secretion in trypsin-treated islets of Langerhans. Horm. Metab. Res.5, 325–329 (1973)
Moskalewski, S., Sochánska, K.R., Wiwatowski, T.: The presence of a factor toxic to pancreatic beta cells in some collagenase preparations. Bull. Acad. Pol.22, 127–130 (1974)
Temin, H.M., Pierson Jr., R.W., Dulak, N.C.: The role of serum in the control of multiplication of avian and mammalian cells. In: Culture, growth, nutrition and metabolism of cells in culture (eds. G.H. Rothblat, V.J. Cristofalo), pp. 49–81. New York: Academic Press 1972
Andersson, A., Gunnarsson, R., Hellerström, C.: Long-term effects of a low-glucose medium on glucose metabolism and insulin biosynthesis and release of isolated pancreatic islets maintained in tissue culture. Acta endocr. (Kbh.)77 (Suppl. 190), 8 (1974)
Howell, S.L., Green, I.C., Montague, W.: A possible role of adenylate cyclase in the long-term dietary regulation of insulin secretion from rat islets of Langerhans. Biochem. J.136, 343–349 (1973)
Ashcroft, S.J.H., Bassett, J.M., Randle, P.J.: Insulin secretion mechanisms and glucose metabolism in isolated islets. Diabetes21 (Suppl. 2), 538–545 (1972)
Ashcroft, S.J.H., Weerasinghe, L.C.C., Randle, P.J.: Interrelationship of islet metabolism, adenosine triphosphate content and insulin release. Biochem. J.132, 223–231 (1973)
Hellman, B., Idahl, L.-Å., Lernmark, Å., Sehlin, J., Täljedal, I.-B.: The pancreatic β-cell recognition of insulin secretagogues. Effects of calcium and sodium on glucose metabolism and insulin release. Biochem. J.138, 33–45 (1974)
Andersson, A., Borglund, E., Brolin, S.: Effects of glucose on the content of ATP and glycogen and the rate of glucose phosphorylation of isolated pancreatic islets maintained in tissue culture. Biochem. biophys. Res. Commun.56, 1045–1051 (1974)
Hellman, B., Idahl, L.-Å., Danielsson, Å.: Adenosine triphosphate levels of mammalian pancreatic B cells after stimulation with glucose and sulfonylureas. Diabetes18, 509–516 (1969)
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Buitrago, A., Gylfe, E., Hellman, B. et al. Function of microdissected pancreatic islets cultured in a chemically defined medium. I. Insulin content and release. Diabetologia 11, 535–540 (1975). https://doi.org/10.1007/BF01222103
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF01222103