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Vol. 83, No. 2, 2006
Issue release date: August 2006
Section title: Original Paper
Neuroendocrinology 2006;83:123–130
(DOI:10.1159/000094875)

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

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Article / Publication Details

First-Page Preview
Abstract of Original Paper

Received: 3/7/2006
Accepted: 4/13/2006
Published online: 8/25/2006

Number of Print Pages: 8
Number of Figures: 3
Number of Tables: 1

ISSN: 0028-3835 (Print)
eISSN: 1423-0194 (Online)

For additional information: http://www.karger.com/NEN

Abstract

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.


  

Author Contacts

Ludwig Wagner
Department of Medicine III, Medical University Vienna
Währinger Gürtel 18–20, AT–1090 Vienna (Austria)
Tel. +43 1 404 004 341, Fax +43 1 404 007 790
E-Mail ludwig.wagner@meduniwien.ac.at

  

Article Information

Received: April 13, 2006
Accepted after revision: July 3, 2006
Published online: August 3, 2006
Number of Print Pages : 8
Number of Figures : 3, Number of Tables : 1, Number of References : 51

  

Publication Details

Neuroendocrinology (International Journal for Basic and Clinical Studies on Neuroendocrine Relationships)

Vol. 83, No. 2, Year 2006 (Cover Date: August 2006)

Journal Editor: Millar, R. (Edinburgh)
ISSN: 0028–3835 (print), 1423–0194 (Online)

For additional information: http://www.karger.com/NEN


Article / Publication Details

First-Page Preview
Abstract of Original Paper

Received: 3/7/2006
Accepted: 4/13/2006
Published online: 8/25/2006

Number of Print Pages: 8
Number of Figures: 3
Number of Tables: 1

ISSN: 0028-3835 (Print)
eISSN: 1423-0194 (Online)

For additional information: http://www.karger.com/NEN


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