Establishment of a protocol to extend the lifespan of human hormone-secreting pituitary adenoma cells
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ENDOCRINE METHODS AND TECHNIQUES
Establishment of a protocol to extend the lifespan of human hormone-secreting pituitary adenoma cells Aurora Aiello1,2 Maria Francesca Cassarino3 Simona Nanni2 Antonella Sesta3,4 Francesco Ferraú5 Claudio Grassi6 Marco Losa7 Francesco Trimarchi8 Alfredo Pontecorvi2 Salvatore Cannavò9 Francesca Pecori Giraldi3,4 Antonella Farsetti 1 ●
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Received: 21 December 2016 / Accepted: 17 April 2017 © Springer Science+Business Media New York 2017
Abstract Purpose The aim of this study was to generate immortalized human anterior pituitary adenoma cells. Reliable cell models for the study of human pituitary adenomas are as yet lacking and studies performed so far used repeated passaging of freshly excised adenomas, with the attendant limitations due to limited survival in culture, early senescence, and poor reproducibility. Methods & Results We devised a technique based upon repeated co-transfections of two retroviral vectors, one carrying the catalytic subunit of human telomerase, hTERT,
the other SV40 large T antigen. This approach extended the lifespan of cells derived from a human growth hormonesecreting adenoma up to 18 months while retaining morphology of primary cells, growth hormone synthesis and growth hormone secretion. Conclusions Our attempt represents the first demonstration of successful lifespan extension of human growth hormonesecreting pituitary adenoma cells via co-transfection of hTERT and SV40T and paves the way to future attempts to obtain stable cell lines. Keywords GH-secreting adenoma Immortalization Pituitary cell line TERT SV40 T-antigen ●
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Aurora Aiello, Maria Francesca Cassarino and Simona Nanni contributed equally to this work.
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* Antonella Farsetti [email protected] 1
Institute of Cell Biology and Neurobiology, National Research Council, Rome 00143, Italy
Introduction
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Institute of Medical Pathology, Università Cattolica, Rome 00168, Italy
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Neuroendocrinology Research Laboratory, Istituto Auxologico Italiano IRCCS, Milan 20095, Italy
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Deparment of Clinical Sciences & Community Health, Università di Milano, Milan 20122, Italy
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Department of Clinical and Experimental Medicine, University of Messina, Messina 98125, Italy
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Institute of Human Physiology, Università Cattolica, Rome 00168, Italy
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Department of Neurosurgery, Ospedale San Raffaele IRCCS, Milan 20132, Italy
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Accademia Peloritana dei Pericolanti, University of Messina, Messina 98125, Italy
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Department of Childhood and Adulthood Human Pathology G. Barresi, University of Messina, Messina 98125, Italy
The study of human pituitary adenomas is hampered by paucity of surgical specimens and by the lack of comparable animal models. Major efforts to obtain cell lines from pituitary tumors have been expended over the years, beginning with immortalization of pituitary cells from experimental pituitary tumors induced by radiation, carcinogens or estrogen treatment [1–3]. Subsequently, transgenic technology allowed the establishment of cell lines der
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