Long-Term in vitro Growth of Human Insulin-Secreting Insulinoma CellsGartner 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
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.
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
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
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