Abstract
Objective: Mifepristone has been recently demonstrated to diminish breakthrough bleeding in users of progestin contraceptives. The purpose of this study was to investigate the use of cultured endometrial explants to study the effects of mifepristone and medroxyprogesterone acetate (MPA) on endometrial sex steroid receptors. Design: Prospective histological, immunocytochemical and molecular analysis. Materials and Methods: Human endometrium was obtained by biopsy during the secretory phase, cut into 1 mm3 pieces, and cultured in D-MEM/F-12 with 5% charcoal-stripped fetal bovine serum on Millicell Organotypic culture inserts for 24 hours. Thereafter, explants were cultured for 48 hours in serum-free media in the absence and presence of 17 beta-estradiol (E2, 1 nm), MPA (100 nm), E2 plus MPA, mifepristone (1 um), and MPA plus mifepristone, respectively. Explants were then frozen for total RNA isolation or fixed in 10% formalin for morphological and immunocytochemical analysis. H & E staining was performed on sections before and after culture. Levels of ER and PR mRNA were determined by RT-PCR while protein levels were evaluated by immunocytochemistry. Results: Explants showed little or no necrosis up to 72 hours of culture on H & E staining. High molecular weight, non-degraded RNA was demonstrated on gel electrophoresis after 72 hours in culture. Using RT-PCR analysis with beta-actin as reference, PR expression was seen to increase after E2, decrease after MPA and slightly increase in the presence of E2 and MPA. These changes were further confirmed by immunocytochemical analysis. It was also demonstrated by immunocytochemistry that MPA decreased PR and ER expression whereas the combination of MPA and mifepristone increased PR and ER expression. Conclusion: Tissue viability of endometrial explants can be maintained in culture up to 72 hours and can be used to directly evaluate the induction or repression of gene products in the endometrium by progestins and anti-progestins. Supported by: NIH HD043189.
Published Version (
Free)
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
