223: Progesterone inhibits cervical tissue formation in a 3D culture system in a dose-dependent fashion

3D culture system in a dose-dependent fashion Michael House, Serkalem Tadesse, Errol Norwitz, David Kaplan Tufts Medical Center, Maternal Fetal Medicine, Boston, MA, Tufts University, Biomedical Engineering, Medford, MA, Yale University, Obstetrics and Gynecology, New Haven, CT OBJECTIVE: Progesterone supplementation can prevent preterm birth in some women with cervical shortening, suggesting that progesterone may directly affect remodeling of cervical tissue. A previously described 3D culture system was used to test progesterone effects on cervical tissue synthesis. STUDY DESIGN: Fibroblasts were obtained from cervical biopsies of non-pregnant premenopausal women undergoing hysterectomy for benign disease. Fibroblasts from passage 5 were seeded on silk sponge scaffolds and cultured in spinner flasks using previously validated culture system (House, Tissue Eng Part A, 2012). Culture media included DMEM 5% charcoal-stripped FBS estradiol 10-8M. Five progesterone conditions were used: 0 (vehicle), 10-9M, 10-8M, 10-7M and 10-7M 10-6M mifepristone. Scaffolds were cultured for 4 weeks and assayed for collagen production (hydroxyproline), histology and immunohistochemistry (IHC). Cervical-like tissue was assayed for estrogen and progesterone receptors using IHC and western blot. Experiments were repeated in triplicate with cells from 3 different women. RESULTS: Cervical cells reliably synthesized cervical-like tissue in 3D. Both progesterone and estrogen receptors were documented by IHC and western blot. A dose-dependent inhibitory effect of progesterone was observed. The highest progesterone concentration (10-7M) was associated with the least amount of collagen synthesis. Collagen synthesis increased progressively as progesterone concentration decreased (p .01). This effect was abrogated by mifepristone (p .05). Differences in tissue morphology on histology and IHC correlated with collagen synthesis. CONCLUSION: Progesterone inhibited the synthesis of cervical-like tissue in 3D culture from cervical fibroblasts in a dose-dependent manner. This effect was abrogated by mifepristone, a progesterone receptor antagonist. This hormonally-responsive in vitro culture system could be used to study the mechanism of progesterone effects on the cervix. 224 Role of (pro)rennin and its receptor in pathogenesis of preeclampsia: a rat model and human patients studies Kelsey Kelso, Russell Fothergill, Steven Allen, Richard Jones, Thomas Kuehl, Mohammad Uddin Scott & White Memorial Hospital / Texas AM placental (pro)renin for NP:152 79 and PreE:302 42 ng/g tissue). In addition to serving as a source of (pro)renin, the placenta is also a site for signaling as ERK1/2 phosphorylation is greater (p 0.05) in placental tissue of preE rats. CONCLUSION: Together with the upregulation of ERK1/2 phosphorylation in placenta of the rat model, there is now evidence of (pro)renin and its receptor associated novel RAS activation to play a role in preE pathogenesis through (pro)renin receptor-mediated detrimental cellular signaling at the placental boundary. This offers an opportunity for interventional treatments with signal inhibitors.