Endometrial cells with high ALDH activity contribute to uterine development and regeneration

Endometrial cells with high ALDH activity contribute to uterine development and regeneration | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Endometrial cells with high ALDH activity contribute to uterine development and regeneration Diana Monsivais, SUNI TANG, Anna Unser, Peixin Jiang, Sydney Parks, and 5 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6831916/v3 This work is licensed under a CC BY 4.0 License Status: Posted Version 3 posted You are reading this latest preprint version Show more versions Abstract Adult stem cells are thought to drive the regenerative potential of the endometrium and contribute to the pathogenesis of endometriosis, however, their identity and defining features remain to be characterized. Here, we used in vivo and in vitro approaches to demonstrate that cells with high aldehyde dehydrogenase 1 activity (ALDH HI cells) were long lived progenitors in the endometrium with a higher organoid formation capacity, long-term passaging potential, and stemness gene signatures. Using lineage tracing with an Aldh1a1 cre/ERT2 ; ROSA26 tdTomato reporter mouse, Aldh1a1 + epithelial cells expanded during postnatal development, Aldh1a1 + stromal cells expanded during estrous cycling, and both populations of Aldh1a1 + cells were present during postpartum repair. In response to ovariectomy or exogenous estradiol, we found that ALDH1A1 + cells localized to glandular crypts of the endometrium or throughout the luminal epithelium, respectively, indicating that their spatial localization is hormone sensitive. Functionally, we found that selective ablation of ALDH1A1 + cells in Aldh1a1 cre/ERT2 ; ROSA26- DTR flox/flox mice decreased endometrial gland number and FOXA2 expression . These findings were recapitulated in the human endometrium, where endometrial epithelial organoids with high ALDH activity (ALDH HI cells) showed a higher organoid formation capacity than ALDH LO cells and displayed unique transcriptomes with fewer luminal-like ciliated cells. Overall, our studies indicate that ALDH1A1 + cells are hormone-sensitive adult stem cells in the endometrium with regenerative potential that are critical for endometrial development and function. Biological sciences/Stem cells/Self-renewal Biological sciences/Developmental biology/Stem-cell niche Lineage tracing organoid endometrium stem cells endometriosis inflammation progenitors Full Text Additional Declarations The authors declare no competing interests. Supplementary Files ALDH1A1SupplementalData5.6.26revisedfinal.pdf SupplementalTableS1MouseOrganoidHivsLov2.xlsx SupplementalTableS1MouseOrganoidHivsLo.xlsx Supplemental Table S1- Mouse Organoid Hi vs. Lo SupplementalTableS5.xlsx Supplemental Table S5. Sample Information SupplementalTableS5revised.xlsx Cite Share Download PDF Status: Posted Version 3 posted You are reading this latest preprint version Show more versions Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-6831916","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":611576445,"identity":"dd4ca05b-373c-474a-be5f-fdad8d4c4ba8","order_by":0,"name":"Diana 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Using lineage tracing with an\u0026nbsp;\u003cem\u003eAldh1a1\u003c/em\u003e\u003csup\u003e\u003cem\u003ecre/ERT2\u003c/em\u003e\u003c/sup\u003e\u003cem\u003e; ROSA26\u003c/em\u003e\u003csup\u003e\u003cem\u003etdTomato\u003c/em\u003e\u003c/sup\u003e\u003cem\u003e\u0026nbsp;\u003c/em\u003ereporter mouse,\u0026nbsp;\u003cem\u003eAldh1a1\u003c/em\u003e\u003csup\u003e+\u0026nbsp;\u003c/sup\u003eepithelial cells expanded during postnatal development,\u0026nbsp;\u003cem\u003eAldh1a1\u003c/em\u003e\u003csup\u003e+\u0026nbsp;\u003c/sup\u003estromal cells expanded during estrous cycling, and both populations of\u0026nbsp;\u003cem\u003eAldh1a1\u003c/em\u003e\u003csup\u003e+\u0026nbsp;\u003c/sup\u003ecells were present during postpartum repair. In response to ovariectomy or exogenous estradiol, we found that ALDH1A1\u003csup\u003e+\u003c/sup\u003e\u0026nbsp;cells localized to glandular crypts of the endometrium or throughout the luminal epithelium, respectively, indicating that their spatial localization is hormone sensitive. Functionally, we found that selective ablation of ALDH1A1\u003csup\u003e+\u003c/sup\u003e\u0026nbsp;cells in\u0026nbsp;\u003cem\u003eAldh1a1\u003c/em\u003e\u003csup\u003e\u003cem\u003ecre/ERT2\u003c/em\u003e\u003c/sup\u003e\u003cem\u003e; ROSA26-\u003c/em\u003eDTR\u003csup\u003eflox/flox\u0026nbsp;\u003c/sup\u003emice decreased endometrial gland number and FOXA2 expression\u003cem\u003e.\u003c/em\u003e\u0026nbsp;These findings were recapitulated in the human endometrium, where endometrial epithelial organoids with high ALDH activity (ALDH\u003csup\u003eHI\u003c/sup\u003e\u0026nbsp;cells) showed a higher organoid formation capacity than ALDH\u003csup\u003eLO\u003c/sup\u003e\u0026nbsp;cells and displayed unique transcriptomes with fewer luminal-like ciliated cells. Overall, our studies indicate that ALDH1A1\u003csup\u003e+\u003c/sup\u003e\u0026nbsp;cells are hormone-sensitive adult stem cells in the endometrium with regenerative potential that are critical for endometrial development and function.\u003c/p\u003e","manuscriptTitle":"Endometrial cells with high ALDH activity contribute to uterine development and regeneration","msid":"","msnumber":"","nonDraftVersions":[{"code":3,"date":"2026-05-26 18:03:28","doi":"10.21203/rs.3.rs-6831916/v3","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}},{"code":2,"date":"2026-03-24 18:56:24","doi":"10.21203/rs.3.rs-6831916/v2","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}},{"code":1,"date":"2025-07-08 10:48:03","doi":"10.21203/rs.3.rs-6831916/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"92569f02-2bca-4a95-98d5-803592864faa","owner":[],"postedDate":"May 26th, 2026","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":67667576,"name":"Biological sciences/Stem cells/Self-renewal"},{"id":67667577,"name":"Biological sciences/Developmental biology/Stem-cell niche"}],"tags":[],"updatedAt":"2026-03-23T15:12:27+00:00","versionOfRecord":[],"versionCreatedAt":"2026-05-26 18:03:28","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v3","identity":"rs-6831916","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-6831916","identity":"rs-6831916","version":["v3"]},"buildId":"WvIrzKhiLBfengagbw6Ux","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}