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Development and Characterization of a Simian Virus 40 Immortalized Bovine Endometrial Stromal Cell Line

Centre Hospitalier Universitaire de Québec, Centre Hospitalier de l’Université Laval, and Centre de Recherche en Biologie de la Reproduction et Département d’Obstétrique et Gynécologie (N.K., P.C., M.A.F.), Département de Physiologie (J.P.T.), Université Laval, Québec, Canada G1K 7P4

Address all correspondence and requests for reprints to: Dr. Michel A. Fortier, Ontogénie et Reproduction, Room T-1-49, Centre Hospitalier Universitaire de Québec (CHUL), 2705 Boulevard Laurier, Québec, G1V 4G2 Québec, Canada. E-mail: MAFortier{at}crchul.ulaval.ca.

In ruminants, interferon- (IFN) is the maternal recognition signal inhibiting prostaglandin (PG) F₂ production by endometrial epithelial cells and stimulating interferon-stimulated genes in the stroma. Stromal cells mediate the action of progesterone on epithelial cells during pregnancy. Our working hypothesis is that IFN acts as a molecular switch that turns on PGE₂ production in endometrial stromal cells while suppressing PGF₂ production from epithelial cells. In this report we document immortalization and functional characterization of a bovine stromal cell line from the caruncular region of the endometrium [caruncular stromal cell (CSC)]. Primary stromal cells were immortalized by nucleofection with simian virus 40 large T antigen and integrase. The resulting cell line, CSC, expresses stromal cell-specific vimentin, estrogen, and progesterone receptors, and is amenable for transient transfection. Basal and stimulated production of PGE₂ is higher than PGF₂ and associated with cyclooxygenase (COX) 2 expression. Phorbol myristate acetate (PMA) and IFN up-regulate COX2 and PG production in a dose-dependent manner. When added together, low concentrations of IFN inhibit PMA-induced COX2 expression; whereas this inhibition is lost at high concentrations. Expression of signal transducer and activator of transcription 1 is induced by IFN at all concentrations studied but is not modulated by PMA. Because expression of signal transducer and activator of transcription 1 does not exhibit the biphasic response to IFN, we investigated the p38 MAPK pathway using the selective inhibitor SB203580. Inhibition of the p38 MAPK pathway abolishes IFN action on PG production. In summary, CSC appears as a good stromal cell model for investigating the molecular mechanisms related to IFN action and PG production in the bovine.