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To mimic the secretory phase, organoids were sequentially treated with 17\u03b2-estradiol\u00a0(E2) and progesterone (P4) and subjected to RNA sequencing. Differentially expressed genes (DEGs) were identified using DESeq2 (lfcThreshold\u2009=\u20090, log<jats:sub>2<\/jats:sub> Fold Change\u2009\u2265\u20091.0 or\u2009\u2264\u2009\u22121.0), and a principal component analysis (PCA) plot was generated. Functional enrichment analysis was performed by overrepresentation analysis and Gene Set Enrichment Analysis (GSEA). To functionally assess proliferation, OrganoSeg surface measurements were performed before (T<jats:sub>0<\/jats:sub>) and after (T<jats:sub>1<\/jats:sub>) differentiation of organoids, and T<jats:sub>1<\/jats:sub>\/T<jats:sub>0<\/jats:sub> ratios were calculated to determine the proliferation rate.<\/jats:p>\n              <\/jats:sec><jats:sec>\n                <jats:title>Results<\/jats:title>\n                <jats:p>Although the PCA plot did not show clear clustering of the fertile and infertile samples, 363 significant DEGs (129 upregulated and 234 downregulated) were detected in infertile compared to fertile organoids. Mainly cell cycle processes were highly enriched in infertile organoids. Thus, we hypothesised that proliferative activity during differentiation may be higher in infertile organoids compared to fertile organoids. However, this could not be validated by cell surface measurements.<\/jats:p>\n              <\/jats:sec><jats:sec>\n                <jats:title>Conclusions<\/jats:title>\n                <jats:p>This study revealed that cell cycle processes were enriched in E2\/P4-treated infertile endometrial organoids as compared to fertile organoids. This could reflect persistently higher proliferative activity of the endometrial epithelial cells in differentiated infertile organoids compared to fertile organoids. To confirm this hypothesis, further studies are warranted.<\/jats:p>\n              <\/jats:sec>","DOI":"10.1007\/s10815-024-03173-y","type":"journal-article","created":{"date-parts":[[2024,7,12]],"date-time":"2024-07-12T14:02:00Z","timestamp":1720792920000},"page":"2405-2418","update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Enrichment of cell cycle pathways in progesterone-treated endometrial organoids of infertile women compared to fertile women"],"prefix":"10.1007","volume":"41","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-3542-6215","authenticated-orcid":false,"given":"B. N.","family":"Bui","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8527-4585","authenticated-orcid":false,"given":"A. 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Written informed consent was obtained from all study participants.","order":2,"name":"Ethics","group":{"name":"EthicsHeading","label":"Ethics approval"}},{"value":"The SCRATCH and SCRaTCH-OFO trials and the current endometrial organoid transcriptome study received funding from public sources and Merck, but none of these sources had input in the study design and analysis. In addition, several authors report having received personal fees for consultations, conference attendance, and travel as follows: BB from Gedeon Richter Benelux and Guerbet; GS from Guerbet; and SM from Abbott Pharmaceuticals, Ferring, Merck, Oxolife and IBSA. FB is a paid member of the advisory boards of Merck and Ferring and also received speakers\u2019 fees from Besins Healthcare, Cooper Surgical and Merck. All of these personal fees were unrelated to the study reported in this manuscript. The remaining authors declare no conflicts of interest relevant to this study.","order":3,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}]}}