A new procedure has been developed for dissociating anterior pituitary tissue and producing a viable suspension of single cells. The procedure involves incubation of small tissue blocks in 1 mg/ml trypsin (15 min), followed by incubation in 8 µg/ml neuraminidase and 1 mM EDTA (15 min), followed by mechanical dispersion. Cell yields are ∼55%, based on recovered DNA. By electron microscopy five types of secretory cells (somatotrophs, mammotrophs, thyrotrophs, gonadotrophs, and corticotrophs) plus endothelial and follicular cells can be identified and are morphologically well preserved up to 20 h after dissociation. Throughout this period, the cells incorporate linearly [3H]leucine into protein for up to 4 h at a rate 90% greater than hemipituitaries, and they synthesize, transport intracellularly, and release the two major pituitary secretory products, growth hormone and prolactin. Immediately after dissociation the cells' ability to respond to secretogogues (high K+ and dibutyryl cyclic AMP) is impaired, but after a 6–12-h culture period, the cells apparently recover and discharge 24% and 52%, respectively, of their content of prelabeled growth hormone over a 3-h period in response to these two secretogogues. This represents a stimulation of 109% and 470% over that released by cells incubated in control medium. The results demonstrate that function and morphologic integrity are preserved in this cell system. Therefore it is suitable for the study of various aspects of pituitary secretion and its control.
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1 November 1973
Article|
November 01 1973
HORMONE SECRETION BY CELLS DISSOCIATED FROM RAT ANTERIOR PITUITARIES
Colin R. Hopkins,
Colin R. Hopkins
From The Rockefeller University, New York 10021.
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Marilyn G. Farquhar
Marilyn G. Farquhar
From The Rockefeller University, New York 10021.
Search for other works by this author on:
Colin R. Hopkins
From The Rockefeller University, New York 10021.
Marilyn G. Farquhar
From The Rockefeller University, New York 10021.
Dr. Hopkins' present address is the Department of Histology and Cell Biology (Medical), The University of Liverpool, England. Dr. Farquhar's present address is the Section of Cell Biology, Yale University School of Medicine, New Haven, Connecticut 06510.
Received:
March 27 1973
Revision Received:
July 10 1973
Online ISSN: 1540-8140
Print ISSN: 0021-9525
Copyright © 1973 by The Rockefeller University Press
1973
J Cell Biol (1973) 59 (2): 276–303.
Article history
Received:
March 27 1973
Revision Received:
July 10 1973
Citation
Colin R. Hopkins, Marilyn G. Farquhar; HORMONE SECRETION BY CELLS DISSOCIATED FROM RAT ANTERIOR PITUITARIES . J Cell Biol 1 November 1973; 59 (2): 276–303. doi: https://doi.org/10.1083/jcb.59.2.276
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