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Long-term culture of human corticotropin-secreting adenomas on extracellular matrix and evaluation of serum-free conditions

Secretory aspects

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Summary

Tissues from 12 human corticotropin-secreting adenomas, obtained during surgery for Cushing's disease (CD, ten cases) or Nelson's Syndrome (NS, two cases), were exclusively mechanically dispersed. Single cells and cell aggregates were plated on extracellular matrix derived from bovine corneal endothelia. Functional responses to physiological stimuli were analyzed by measuring human β-endorphin (βh-EP) immunoreactivity (IR) by radioimmunoassay in the culture medium. All adenomas responded with stimulated secretory activity to arginine vasopressin (AVP), corticotropin-releasing factor (CRF), or both. Cortisol higher than 10−8 M suppressed basal secretion and CRF- or AVP-stimulated βh-EP-IR secretion. There was no consistent difference in response of the cells when cultured in medium containing 10% fetal calf serum (FCS) or in serum-free conditions. A change of cells from serum to serum-free conditions usually resulted in 10%–50% reduction in the basal secretion level that remained stable for at least 2 weeks and, in one case (NS), 10 weeks. In cells maintained in medium supplemented with 5% serum obtained from the respective patients 40 min after adenoma removal, basal secretion was suppressed to 60% of the baseline level in a 10% FCS control. Long-term incubation with CRF (10−9 M) showed sustained stimulation of hormone secretion. No remarkable cell proliferation was observed under basal conditions or during long-term, low-dose incubation with cholera toxin (10−12 M) in two cases (CD), or CRF (10−9 M) in two cases (NS, CD). Parallel β-EP-IR and adrenocorticotropin secretion was verified in selected cases.

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Author notes

  1. M. Westphal

    Present address: Neurologische Klinik, Neurochirurgische Abteilung, Universitäts-Krankenhaus Eppendorf, Martinistrasse 52, D-2000, Hamburg 20, Germany

  2. P. Jaquet

    Present address: Clinique Endocrinologique, Hôpital de la Conception, Rue Saint Pierre, F-13385, Marseille, France

Authors and Affiliations

  1. Laboratory of Molecular Endocrinology, School of Medicine, University of California, 94143, San Francisco, CA, USA

    M. Westphal, P. Jaquet & C. B. Wilson

  2. Departments of Gynecology and Reproductive Sciences, and Neurological Surgery, School of Medicine, University of California, 94143, San Francisco, CA, USA

    M. Westphal, P. Jaquet & C. B. Wilson

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  1. M. Westphal
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Additional information

Supported by Grants GMO 02907, NIH-AM-06097, and NIDA DA-03434 from the National Institutes of Health, USA. The work has been submitted in abstract form to the XV Acta Endocrinologica Congress, Helsinki, Finland. M. W. is recipient of a research training grant from the Deutsche Forschungsgemeinschaft. P. J. was supported by INSERM grant CRL-80-40-19, NIH-HD-08035.

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Westphal, M., Jaquet, P. & Wilson, C.B. Long-term culture of human corticotropin-secreting adenomas on extracellular matrix and evaluation of serum-free conditions. Acta Neuropathol 71, 111–118 (1986). https://doi.org/10.1007/BF00687971

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  • DOI: https://doi.org/10.1007/BF00687971

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