We report the establishment and preliminary characterization of a stable steroidogenic granulosa cell line, JC-410. This cell line was obtained by spontaneous immortalization of a primary culture of porcine granulosa cells. Cultured JC-410 cells produced less progesterone than granulosa cells in primary culture. Progesterone synthesis by JC-410 cells was approximately 10% and 1% of the amount produced by granulosa cells from small and medium sized follicles, respectively. Although FSH and LH did not change progesterone levels in cultured JC-410 cells, forskolin and cholera toxin induced a 2.6- and 2.75-fold increase, respectively, versus control. The JC-410 cells responded to 0.1, 1 and 5 mM cAMP with an increase in progesterone synthesis of 2.5-, 28- and 49-fold versus control, respectively, after a 24 h incubation. No detectable levels of estradiol-17beta were found in JC-410 cells after 48 h in culture. However, addition of 0.01, 0.1 and 1 microM androstenedione elevated the levels of estradiol-17beta to 0.028, 0.3 and 1.21 pg/microg protein, respectively. The level of expression of 3betaHSD, aromatase and P450scc genes in JC-410 cells is of similar magnitude to the level of expression in granulosa cells in primary culture. The JC410 cells have been maintained in culture for more than one year during which their population doubled over 100 times. We conclude that JC-410 is a stable cell line that lost responsiveness to the gonadotropins during the process of immortalization, but retained its steroid biosynthetic capability and the expression of key steroidogenic genes. These characteristics may reflect features of cells arrested in an early stage of granulosa cell differentiation.

This article has been cited by other articles:

				M. Ghadami, S.A. Salama, N. Khatoon, R. Chilvers, M. Nagamani, P.J. Chedrese, and A. Al-Hendy Toward gene therapy of primary ovarian failure: adenovirus expressing human FSH receptor corrects the Finnish C566T mutation Mol. Hum. Reprod., January 1, 2008; 14(1): 9 - 15. [Abstract] [Full Text] [PDF]

				S. D. McKenna, M. Pietropaolo, E. G. Tos, A. Clark, D. Fischer, D. Kagan, B. Bao, P. J. Chedrese, and S. Palmer Pharmacological Inhibition of Phosphodiesterase 4 Triggers Ovulation in Follicle-Stimulating Hormone-Primed Rats Endocrinology, January 1, 2005; 146(1): 208 - 214. [Abstract] [Full Text] [PDF]

				R. J. Urban, Y. H. Bodenburg, J. Jiang, L. Denner, and J. Chedrese Protein kinase C{iota} enhances the transcriptional activity of the porcine P-450 side-chain cleavage insulin-like response element Am J Physiol Endocrinol Metab, June 1, 2004; 286(6): E975 - E979. [Abstract] [Full Text] [PDF]

				M. C. Henson and P. J. Chedrese Endocrine Disruption by Cadmium, a Common Environmental Toxicant with Paradoxical Effects on Reproduction Experimental Biology and Medicine, May 1, 2004; 229(5): 383 - 392. [Abstract] [Full Text] [PDF]

				A. D. Smida, X. P. Valderrama, M. C. Agostini, M. A. Furlan, and J. Chedrese Cadmium Stimulates Transcription of the Cytochrome P450 Side Chain Cleavage Gene in Genetically Modified Stable Porcine Granulosa Cells Biol Reprod, January 1, 2004; 70(1): 25 - 31. [Abstract] [Full Text] [PDF]

				W. Xing and M. R. Sairam Retinoic Acid Mediates Transcriptional Repression of Ovine Follicle-Stimulating Hormone Receptor Gene via a Pleiotropic Nuclear Receptor Response Element Biol Reprod, July 1, 2002; 67(1): 204 - 211. [Abstract] [Full Text] [PDF]

				W. Xing, N. Danilovich, and M. R. Sairam Orphan Receptor Chicken Ovalbumin Upstream Promoter Transcription Factors Inhibit Steroid Factor-1, Upstream Stimulatory Factor, and Activator Protein-1 Activation of Ovine Follicle-Stimulating Hormone Receptor Expression via Composite cis-Elements Biol Reprod, June 1, 2002; 66(6): 1656 - 1666. [Abstract] [Full Text] [PDF]

				C.L. Swan, M.C. Agostini, P.M. Bartlewski, V. Feyles, R.J. Urban, and P.J. Chedrese Effects of Progestins on Progesterone Synthesis in a Stable Porcine Granulosa Cell Line: Control of Transcriptional Activity of the Cytochrome P450 Side-Chain Cleavage Gene Biol Reprod, April 1, 2002; 66(4): 959 - 965. [Abstract] [Full Text] [PDF]

				W. Xing and M. R. Sairam Role of CACC-Box in the Regulation of Ovine Follicle-Stimulating Hormone Receptor Expression Biol Reprod, October 1, 2001; 65(4): 1142 - 1149. [Abstract] [Full Text] [PDF]

				W. Xing and M. Ram Sairam Characterization of Regulatory Elements of Ovine Follicle-Stimulating Hormone (FSH) Receptor Gene: The Role of E-Box in the Regulation of Ovine FSH Receptor Expression Biol Reprod, February 1, 2001; 64(2): 579 - 589. [Abstract] [Full Text]

				Y. Nishi, T. Yanase, Y.-M. Mu, K. Oba, I. Ichino, M. Saito, M. Nomura, C. Mukasa, T. Okabe, K. Goto, et al. Establishment and Characterization of a Steroidogenic Human Granulosa-Like Tumor Cell Line, KGN, That Expresses Functional Follicle-Stimulating Hormone Receptor Endocrinology, January 1, 2001; 142(1): 437 - 445. [Abstract] [Full Text] [PDF]

				R. M. Touyz, L. Jiang, and M. Ram Sairam Follicle-Stimulating Hormone Mediated Calcium Signaling by the Alternatively Spliced Growth Factor Type I Receptor Biol Reprod, April 1, 2000; 62(4): 1067 - 1074. [Abstract] [Full Text]

				H. Zhang, M. Vollmer, M. De Geyter, Y. Litzistorf, A. Ladewig, M. Durrenberger, R. Guggenheim, P. Miny, W. Holzgreve, and C. De Geyter Characterization of an immortalized human granulosa cell line (COV434) Mol. Hum. Reprod., February 1, 2000; 6(2): 146 - 153. [Abstract] [Full Text] [PDF]

				N.K. Crellin, M.R. Rodway, C.L. Swan, C. Gillio-Meina, and P.J. Chedrese Dichlorodiphenyldichloroethylene Potentiates the Effect of Protein Kinase A Pathway Activators on Progesterone Synthesis in Cultured Porcine Granulosa Cells Biol Reprod, October 1, 1999; 61(4): 1099 - 1103. [Abstract] [Full Text]

				P. S. Babu, H. Krishnamurthy, P. J. Chedrese, and M. R. Sairam Activation of Extracellular-regulated Kinase Pathways in Ovarian Granulosa Cells by the Novel Growth Factor Type 1 Follicle-stimulating Hormone Receptor. ROLE IN HORMONE SIGNALING AND CELL PROLIFERATION J. Biol. Chem., September 1, 2000; 275(36): 27615 - 27626. [Abstract] [Full Text] [PDF]