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Organoids were treated for 14 days with physiologic levels of estradiol and testosterone to mimic a normal follicular phase or PCOS hormone profiles. Organoids were harvested for immunostaining and ribonucleic acid sequencing.<\/jats:p><\/jats:sec><jats:sec><jats:title>Setting<\/jats:title><jats:p>Academic institution.<\/jats:p><\/jats:sec><jats:sec><jats:title>Patients<\/jats:title><jats:p>Endometrial tissues from 10 premenopausal women undergoing hysterectomy for benign pathologies were obtained following written consent.<\/jats:p><\/jats:sec><jats:sec><jats:title>Main Outcome Measures<\/jats:title><jats:p>Organoid architecture, cell specific markers, functional markers, proliferation, and gene expression were measured.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>A method to generate scaffold-free endometrial organoids containing epithelial and stromal cells was established. These organoids exhibited distinct organization with epithelial cells lining the outer surface and stromal cells in the center of the organoids. Epithelial cells were polarized, organoids expressed cell type specific and functional markers, as well as androgen, estrogen, and progesterone receptors. Treatment with PCOS hormones increased cell proliferation and dysregulated genes in endometrial organoids.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions<\/jats:title><jats:p>A new multicellular, scaffold-free endometrial organoid system was established that resembled physiology of the native endometrium. Excess androgens in PCOS promoted cell proliferation in endometrial organoids, revealing new mechanisms of PCOS-associated with risk of endometrial neoplasia.<\/jats:p><\/jats:sec>","DOI":"10.1210\/clinem\/dgz100","type":"journal-article","created":{"date-parts":[[2019,10,15]],"date-time":"2019-10-15T23:09:15Z","timestamp":1571180955000},"page":"769-780","source":"Crossref","is-referenced-by-count":117,"title":["Scaffold-Free Endometrial Organoids Respond to Excess Androgens Associated With Polycystic Ovarian Syndrome"],"prefix":"10.1210","volume":"105","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-8785-3895","authenticated-orcid":false,"given":"Teerawat","family":"Wiwatpanit","sequence":"first","affiliation":[{"name":"Department of Obstetrics and Gynecology, Feinberg School of Medicine, Northwestern University, Chicago, IL, US"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alina R","family":"Murphy","sequence":"additional","affiliation":[{"name":"Department of 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