Journal Mobile Options
Table of Contents
Vol. 83, No. 2, 2006
Issue release date: August 2006
Neuroendocrinology 2006;83:123–130

Long-Term in vitro Growth of Human Insulin-Secreting Insulinoma Cells

Gartner W. · Koc F. · Nabokikh A. · Daneva T. · Niederle B. · Luger A. · Wagner L.
aDepartment of Medicine III, Medical University Vienna, Vienna, Austria; bAcademy of Science, Sofia, Bulgaria; cDepartment of Endocrine Surgery, and dDivision of Endocrinology and Metabolism, Medical University Vienna, Vienna, Austria

Individual Users: Register with Karger Login Information

Please create your User ID & Password

Contact Information

I have read the Karger Terms and Conditions and agree.

To view the fulltext, please log in

To view the pdf, please log in


Objective: Long-term in vitro maintenance of human insulin-secreting insulinoma cells. Methods: (1) Cell culture of ex vivo-derived insulinoma cell suspensions from 8 individual human donors, using various cell culture medium supplementations; (2) determination of insulin synthesis and secretion using immunocytochemistry and insulin and pro-insulin radioimmunoassays; (3) nestin-immunostaining of long-term in vitro grown insulinoma cell suspensions, and (4) phase-contrast light microscopy for analyzing the in vitro growth characteristics of the insulinoma cells. Results: (1) Parallel persistence of in vitro insulinoma cell proliferation as well as insulin-synthesizing and -secreting capacity depended on both the co-culture of insulinoma cells with human fibroblasts and the supplementation of the cell culture medium with tissue culture supernatant derived from the rodent pituitary adenoma cell line GH-3; (2) immunostaining for insulin and secretagogin confirmed the neuroendocrine origin of the insulinoma cells grown in vitro; (3) insulin secretion capability persisted up to an observation period of 25 weeks; (4) insulin secretion rates after 6 weeks of in vitro growth ranged from 3.5 to 83.3 µU/ml/h/60,000 cells plated, and (5) after long-term in vitro growth of insulinoma-derived cell suspensions with persistent insulin-secreting capacity, nestin staining was observed predominantly in co-cultured fibroblasts. Conclusion: Our data describe for the first time the long-term in vitro culture of insulin-secreting human insulinomas and highlight the importance of β-cell trophic factors for insulinoma cell growth.

Copyright / Drug Dosage

Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher or, in the case of photocopying, direct payment of a specified fee to the Copyright Clearance Center.
Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in goverment regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug.
Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.


  1. Bonner-Weir S, Weir GC: New sources of pancreatic beta-cells. Nat Biotechnol 2005;23:857–861.
  2. Hussain MA, Theise ND: Stem-cell therapy for diabetes mellitus. Lancet 2004;364:203–205.
  3. Gershengorn MC, Hardikar AA, Wei C, Geras-Raaka E, Marcus-Samuels B, Raaka BM: Epithelial-to-mesenchymal transition generates proliferative human islet precursor cells. Science 2004;306:2261–2264.
  4. Dor Y, Brown J, Martinez OI, Melton DA: Adult pancreatic beta-cells are formed by self-duplication rather than stem-cell differentiation. Nature 2004;429:41–46.
  5. Trucco M: Regeneration of the pancreatic beta cell. J Clin Invest 2005;115:5–12.
  6. Murray HE, Paget MB, Downing R: Preservation of glucose responsiveness in human islets maintained in a rotational cell culture system. Mol Cell Endocrinol 2005;238:39–49.
  7. de la Tour D, Halvorsen T, Demeterco C, Tyrberg B, Itkin-Ansari P, Loy M, Yoo SJ, Hao E, Bossie S, Levine F: Beta-cell differentiation from a human pancreatic cell line in vitro and in vivo. Mol Endocrinol 2001;15:476–483.
  8. Abraham EJ, Leech CA, Lin JC, Zulewski H, Habener JF: Insulinotropic hormone glucagon-like peptide-1 differentiation of human pancreatic islet-derived progenitor cells into insulin-producing cells. Endocrinology 2002;143:3152–3161.
  9. Gao R, Ustinov J, Pulkkinen MA, Lundin K, Korsgren O, Otonkoski T: Characterization of endocrine progenitor cells and critical factors for their differentiation in human adult pancreatic cell culture. Diabetes 2003;52:2007–2015.
  10. Shinozuka Y, Okada M, Oki T, Sagane K, Mizui Y, Tanaka I, Katayama K, Murakami-Murofushi K: Altered expression of HES-1, BETA2/NeuroD, and PDX-1 is involved in impaired insulin synthesis induced by glucocorticoids in HIT-T15 cells. Biochem Biophys Res Commun 2001;287:229–235.
  11. Chick WL: Beta cell replication in rat pancreatic monolayer cultures. Effects of glucose, tolbutamide, glucocorticoid, growth hormone and glucagon. Diabetes 1973;22:687–693.
  12. Logsdon CD, Moessner J, Williams JA, Goldfine ID: Glucocorticoids increase amylase mRNA levels, secretory organelles, and secretion in pancreatic acinar AR42J cells. J Cell Biol 1985;100:1200–1208.
  13. Rall L, Pictet R, Githens S, Rutter WJ: Glucocorticoids modulate the in vitro development of the embryonic rat pancreas. J Cell Biol 1977;75:398–409.
  14. Brubaker PL, Drucker DJ: Minireview: glucagon-like peptides regulate cell proliferation and apoptosis in the pancreas, gut, and central nervous system. Endocrinology 2004;145:2653–2659.
  15. Ogata T, Dunbar AJ, Yamamoto Y, Tanaka Y, Seno M, Kojima I: Betacellulin-delta4, a novel differentiation factor for pancreatic beta-cells, ameliorates glucose intolerance in streptozotocin-treated rats. Endocrinology 2005;146:4673–4681.
  16. Suarez-Pinzon WL, Lakey JR, Brand SJ, Rabinovitch A: Combination therapy with epidermal growth factor and gastrin induces neogenesis of human islet (beta)-cells from pancreatic duct cells and an increase in functional beta-cell mass. J Clin Endocrinol Metab 2005;90:3401–3409.
  17. Jensen J, Galsgaard ED, Karlsen AE, Lee YC, Nielsen JH: STAT5 activation by human GH protects insulin-producing cells against interleukin-1beta, interferon-gamma and tumour necrosis factor-alpha-induced apoptosis independent of nitric oxide production. J Endocrinol 2005;187:25–36.
  18. Nielsen JH, Galsgaard ED, Moldrup A, Friedrichsen BN, Billestrup N, Hansen JA, Lee YC, Carlsson C: Regulation of beta-cell mass by hormones and growth factors. Diabetes 2001;50(suppl 1):S25–S29.

    External Resources

  19. Service GJ, Thompson GB, Service FJ, Andrews JC, Collazo-Clavell ML, Lloyd RV: Hyperinsulinemic hypoglycemia with nesidioblastosis after gastric-bypass surgery. N Engl J Med 2005;353:249–254.
  20. Pelengaris S, Khan M: Oncogenic co-operation in beta-cell tumorigenesis. Endocr Relat Cancer 2001;8:307–314.
  21. Creutzfeldt W, Creutzfeldt C, Frerichs H, Track NS, Arnold R: Histochemistry, ultrastructure and hormone content of human insulinomas. Horm Metab Res 1976;suppl 6:7–18.
  22. Flatt PR: Defective regulation of insulin secretion in diabetes and insulinoma. Biochem Soc Trans 1990;18:124–127.
  23. Minn AH, Kayton M, Lorang D, Hoffmann SC, Harlan DM, Libutti SK, Shalev A: Insulinomas and expression of an insulin splice variant. Lancet 2004;363:363–367.
  24. Cavallo MG, Dotta F, Monetini L, Dionisi S, Previti M, Valente L, Toto A, Di Mario U, Pozzilli P: Beta-cell markers and autoantigen expression by a human insulinoma cell line: similarities to native beta cells. J Endocrinol 1996;150:113–120.
  25. Baroni MG, Cavallo MG, Mark M, Monetini L, Stoehrer B, Pozzilli P: Beta-cell gene expression and functional characterisation of the human insulinoma cell line CM. J Endocrinol 1999;161:59–68.
  26. Wagner L, Templ E, Reining G, Base W, Weissel M, Nowotny P, Kaserer K, Waldhausl W: Culture of human insulinoma cells: development of a neuroendocrine tumor cell- and human pancreatic islet cell-specific monoclonal antibody. J Endocrinol 1998;156:469–476.
  27. Macfarlane WM, Cragg H, Docherty HM, Read ML, James RF, Aynsley-Green A, Docherty K: Impaired expression of transcription factor IUF1 in a pancreatic beta-cell line derived from a patient with persistent hyperinsulinaemic hypoglycaemia of infancy (nesidioblastosis). FEBS Lett 1997;413:304–308.
  28. Aynsley-Green A, Polak JM, Bloom SR, Gough MH, Keeling J, Ashcroft SJ, Turner RC, Baum JD: Nesidioblastosis of the pancreas: definition of the syndrome and the management of the severe neonatal hyperinsulinaemic hypoglycaemia. Arch Dis Child 1981;56:496–508.
  29. Macfarlane WM, O’Brien RE, Barnes PD, Shepherd RM, Cosgrove KE, Lindley KJ, Aynsley-Green A, James RF, Docherty K, Dunne MJ: Sulfonylurea receptor 1 and Kir6.2 expression in the novel human insulin-secreting cell line NES2Y. Diabetes 2000;49:953–960.
  30. Macfarlane WM, Shepherd RM, Cosgrove KE, James RF, Dunne MJ, Docherty K: Glucose modulation of insulin mRNA levels is dependent on transcription factor PDX-1 and occurs independently of changes in intracellular Ca2+. Diabetes 2000;49:418–423.
  31. Hohmeier HE, Newgard CB: Cell lines derived from pancreatic islets. Mol Cell Endocrinol 2004;228:121–128.
  32. Hohmeier HE, Mulder H, Chen G, Henkel-Rieger R, Prentki M, Newgard CB: Isolation of INS-1-derived cell lines with robust ATP-sensitive K+ channel-dependent and -independent glucose-stimulated insulin secretion. Diabetes 2000;49:424–430.
  33. Newgard CB, Clark S, BeltrandelRio H, Hohmeier HE, Quaade C, Normington K: Engineered cell lines for insulin replacement in diabetes: current status and future prospects. Diabetologia 1997;40(suppl 2):S42–S47.

    External Resources

  34. Efrat S: Development of engineered pancreatic beta-cell lines for cell therapy of diabetes. Adv Drug Deliv Rev 1998;33:45–52.
  35. Liu HK, Green BD, McClenaghan NH, McCluskey JT, Flatt PR: Deleterious effects of supplementation with dehydroepiandrosterone sulphate or dexamethasone on rat insulin-secreting cells under in vitro culture condition. Biosci Rep 2006;26:31–38.
  36. Lechner A, Nolan AL, Blacken RA, Habener JF: Redifferentiation of insulin-secreting cells after in vitro expansion of adult human pancreatic islet tissue. Biochem Biophys Res Commun 2005;327:581–588.
  37. Beattie GM, Montgomery AM, Lopez AD, Hao E, Perez B, Just ML, Lakey JR, Hart ME, Hayek A: A novel approach to increase human islet cell mass while preserving beta-cell function. Diabetes 2002;51:3435–3439.
  38. Wagner L, Oliyarnyk O, Gartner W, Nowotny P, Groeger M, Kaserer K, Waldhausl W, Pasternack MS: Cloning and expression of secretagogin, a novel neuroendocrine- and pancreatic islet of Langerhans-specific Ca2+-binding protein. J Biol Chem 2000;275:24740–24751.
  39. Wang XC, Xu SY, Wu XY, Song HD, Mao YF, Fan HY, Yu F, Mou B, Gu YY, Xu LQ, Zhou XO, Chen Z, Chen JL, Hu RM: Gene expression profiling in human insulinoma tissue: genes involved in the insulin secretion pathway and cloning of novel full-length cDNAs. Endocr Relat Cancer 2004;11:295–303.
  40. Birkenkamp-Demtroder K, Wagner L, Brandt Sorensen F, Bording Astrup L, Gartner W, Scherubl H, Heine B, Christiansen P, Orntoft TF: Secretagogin is a novel marker for neuroendocrine differentiation. Neuroendocrinology 2005;82:121–138.
  41. Maria-Engler SS, Correa-Giannella ML, Labriola L, Krogh K, Colin C, Lojudice FH, Aita CA, de Oliveira EM, Correa TC, da Silva IC, Genzini T, de Miranda MP, Noronha IL, Vilela L, Coimbra CN, Mortara RA, Guia MM, Eliaschewitz FG, Sogayar MC: Co-localization of nestin and insulin and expression of islet cell markers in long-term human pancreatic nestin-positive cell cultures. J Endocrinol 2004;183:455–467.
  42. Hunziker E, Stein M: Nestin-expressing cells in the pancreatic islets of Langerhans. Biochem Biophys Res Commun 2000;271:116–119.
  43. Zulewski H, Abraham EJ, Gerlach MJ, Daniel PB, Moritz W, Muller B, Vallejo M, Thomas MK, Habener JF: Multipotential nestin-positive stem cells isolated from adult pancreatic islets differentiate ex vivo into pancreatic endocrine, exocrine, and hepatic phenotypes. Diabetes 2001;50:521–533.
  44. Humphrey RK, Bucay N, Beattie GM, Lopez A, Messam CA, Cirulli V, Hayek A: Characterization and isolation of promoter-defined nestin-positive cells from the human fetal pancreas. Diabetes 2003;52:2519–2525.
  45. Selander L, Edlund H: Nestin is expressed in mesenchymal and not epithelial cells of the developing mouse pancreas. Mech Dev 2002;113:189–192.
  46. Ooi GT, Tawadros N, Escalona RM: Pituitary cell lines and their endocrine applications. Mol Cell Endocrinol 2004;228:1–21.
  47. Drucker DJ: Minireview: the glucagon-like peptides. Endocrinology 2001;142:521–527.
  48. Friedrichsen BN, Galsgaard ED, Nielsen JH, Moldrup A: Growth hormone- and prolactin-induced proliferation of insulinoma cells, INS-1, depends on activation of STAT5 (signal transducer and activator of transcription 5). Mol Endocrinol 2001;15:136–148.
  49. Liu JL, Coschigano KT, Robertson K, Lipsett M, Guo Y, Kopchick JJ, Kumar U, Liu YL: Disruption of growth hormone receptor gene causes diminished pancreatic islet size and increased insulin sensitivity in mice. Am J Physiol Endocrinol Metab 2004;287:E405–413.
  50. Asfari M, De W, Postel-Vinay MC, Czernichow P: Expression and regulation of growth hormone (GH) and prolactin (PRL) receptors in a rat insulin producing cell line (INS-1). Mol Cell Endocrinol 1995;107:209–214.
  51. Rajagopal J, Anderson WJ, Kume S, Martinez OI, Melton DA: Insulin staining of ES cell progeny from insulin uptake. Science 2003;299:363.

Pay-per-View Options
Direct payment This item at the regular price: USD 38.00
Payment from account With a Karger Pay-per-View account (down payment USD 150) you profit from a special rate for this and other single items.
This item at the discounted price: USD 26.50