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Biology of Reproduction, Vol 56, 608-616, Copyright © 1997 by Society for the Study of Reproduction
ARTICLES |
CG Gutierrez, BK Campbell and R Webb
Roslin Institute, Roslin, Midlothian, United Kingdom. carlos.gutierrez@bbsrc.ac.uk
The objectives of this study were to develop a serum-free bovine granulosa cell culture system in which FSH-responsive estradiol production could be induced and maintained, and to use this system to evaluate the effects of FSH, insulin, and IGF-I on steroidogenesis and proliferation of bovine granulosa cells from different follicle size categories (< 4-, 4-8, and > 8-mm diameter). In the presence of FSH, granulosa cells from small follicles differentiated in vitro, and estradiol secretion increased with time (p < 0.01) so that by the end of the culture period it was similar to that of cells from large follicles. Granulosa cells from medium and large follicles secreted estradiol throughout the culture period. Cells cultured in plasma- coated culture wells had an increased proliferative response but had lower estradiol production compared to cells cultured under serum-free conditions (p < 0.01). Insulin promoted proliferation and estradiol production by granulosa cells from the three follicle-size categories (p < 0.01). Physiological concentrations of FSH induced proliferation and estradiol secretion (p < 0.01) by granulosa cells in a dose- responsive manner. The inclusion of IGF-I in the culture system enhanced proliferation and estradiol production (p < 0.01), even in the absence of gonadotropic support, demonstrating the gonadotropic characteristics of this growth factor. These results demonstrate the development of a relevant physiological culture system for bovine granulosa cells. This system will permit the detailed study of the key factors controlling the differentiation and proliferation of bovine granulosa cells.
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J. Manuel Silva and C. A. Price Effect of Follicle-Stimulating Hormone on Steroid Secretion and Messenger Ribonucleic Acids Encoding Cytochromes P450 Aromatase and Cholesterol Side-Chain Cleavage in Bovine Granulosa Cells In Vitro Biol Reprod, January 1, 2000; 62(1): 186 - 191. [Abstract] [Full Text]
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T. C. Lavranos, J. M. Mathis, S. E. Latham, B. Kalionis, J. W. Shay, and R. J. Rodgers Evidence for Ovarian Granulosa Stem Cells: Telomerase Activity and Localization of the Telomerase Ribonucleic Acid Component in Bovine Ovarian Follicles Biol Reprod, August 1, 1999; 61(2): 358 - 366. [Abstract] [Full Text]
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K. E. Latham, F. D. M. Bautista, Y. Hirao, M. J. O'Brien, and J. J. Eppig Comparison of Protein Synthesis Patterns in Mouse Cumulus Cells and Mural Granulosa Cells: Effects of Follicle-Stimulating Hormone and Insulin on Granulosa Cell Differentiation In Vitro Biol Reprod, August 1, 1999; 61(2): 482 - 492. [Abstract] [Full Text]
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J. J. Eppig, M. J. O'Brien, F. L. Pendola, and S. Watanabe Factors Affecting the Developmental Competence of Mouse Oocytes Grown In Vitro: Follicle-Stimulating Hormone and Insulin Biol Reprod, December 1, 1998; 59(6): 1445 - 1453. [Abstract] [Full Text]
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D. G. Armstrong, G. Baxter, C. G. Gutierrez, C. O. Hogg, A. L. Glazyrin, B. K. Campbell, T. A. Bramley, and R. Webb Insulin-Like Growth Factor Binding Protein -2 and -4 Messenger Ribonucleic Acid Expression in Bovine Ovarian Follicles: Effect of Gonadotropins and Developmental Status Endocrinology, April 1, 1998; 139(4): 2146 - 2154. [Abstract] [Full Text] [PDF]
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