A Serum-Free System for Primary Cultures of Human Pituitary Adenomas |
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Authors: | Thodou Eleni Ramyar Lily Cohen Arthur I Singer William Asa Sylvia L |
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Institution: | (1) Department of Pathology, Mount Sinai Hospital, 600 University Avenue, M5G 1X5 Toronto, Ontario;(2) Department of Medicine, St. Michael’s Hospital, University of Toronto, Ontario, Canada |
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Abstract: | We report the successful use of a serum-free culture system for primary cultures of human pituitary adenomas. The system utilizes
histiotypic suspension culture with low protein-binding membrane inserts that enable cells to retain their three-dimensional
tissue configuration, closely mimicking the growth pattern in vivo. A serum-free defined medium was developed with CMRL-1969
(Connaught, Willowdale, Ontario, Canada) supplemented with 0.375% albumin bovine Fraction V, 5 μg/mL insulin, 5 μg/mL transferrin,
5 ng/mL sodium selenite, 30 μg/mL putrescine, 6.85 × 10−11
M hydrocortisone, and 3.7 × 10−11
M tri-iodothyronine (T3).
We analyzed eight surgically resected human pituitary adenomas. Basal pituitary hormone secretion measured by radioimmunoassay
of pituitary hormones was compared with hormone hypersecretion in vivo and with control cells of the same tumors cultured
in CMRL-1969 with 10% fetal calf serum. The light microscopic, immunocytochemical, and ultrastructural morphology of cells
cultured in this serum-free histiotypic system was compared with cells cultured in serum-supplemented media and with cells
cultured on collagen-coated plastic; all cultured cells were compared with the morphology of surgically resected tissues of
the same specimens. Basal pituitary hormone secretion during 24-hour incubations correlated with the clinical patterns of
hormone excess; the data were similar in serum-enriched and serum-free cultures, however, hormone secretion decreased less
rapidly in the serum-free cultures. Cells maintained in the histiotypic culture system closely resembled the corresponding
surgically resected tumor using the morphologic parameters and were better preserved than those plated in collagen-coated
plastic wells.
This comparative study indicates that this serum-free histiotypic culture system provides an ideal method of examining pituitary
adenomas in vitro without altering the profile of hormone secretion and cell morphology documented in vivo. This system can
be used to examine the production and effects of a wide range of hormones and growth factors that have been implicated as
causative agents in pituitary tumorigenesis. |
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Keywords: | Pituitary adenohypophysis cell culture radioimmunoassay morphology |
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