{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,13]],"date-time":"2026-10-13T16:20:34Z","timestamp":1791908434914,"version":"4.3.4"},"reference-count":30,"publisher":"Oxford University Press (OUP)","issue":"4","license":[{"start":{"date-parts":[[2025,10,15]],"date-time":"2025-10-15T00:00:00Z","timestamp":1760486400000},"content-version":"vor","delay-in-days":287,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100002301","name":"Estonian Research Council","doi-asserted-by":"publisher","award":["PRG1076"],"award-info":[{"award-number":["PRG1076"]}],"id":[{"id":"10.13039\/501100002301","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004359","name":"Swedish Research Council","doi-asserted-by":"publisher","award":["2024-02530"],"award-info":[{"award-number":["2024-02530"]}],"id":[{"id":"10.13039\/501100004359","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2025,9,22]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:sec>\n                    <jats:title>STUDY QUESTION<\/jats:title>\n                    <jats:p>Can menstrual blood-derived organoids (MB-organoids) and human endometrial stromal cells (MB-ESCs) serve as a physiologically relevant, non-invasive model for studying endometrial function and hormonal response?<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>SUMMARY ANSWER<\/jats:title>\n                    <jats:p>MB-organoids and ESCs recapitulate key structural and functional features of the endometrium, responding to hormonal stimulation in a manner comparable to biopsy-derived models, supporting their use in reproductive research.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>WHAT IS KNOWN ALREADY<\/jats:title>\n                    <jats:p>Endometrial organoids derived from biopsy samples have provided valuable insights into endometrial physiology and implantation. However, their reliance on invasive tissue sampling limits their clinical and research applications. Menstrual blood contains viable endometrial cells, yet its potential for generating functional three-dimensional (3D) models remains underexplored.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>STUDY DESIGN, SIZE, DURATION<\/jats:title>\n                    <jats:p>This cross-sectional, in vitro cell culture study established and characterized 3D-organoids and ESCs derived from menstrual blood, assessing their structural and functional properties as well as their response to hormonal stimulation over a culture period of several weeks. The work was carried out between October 2023 and December 2024, in two European University hospitals.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>PARTICIPANTS\/MATERIALS, SETTING, METHODS<\/jats:title>\n                    <jats:p>Menstrual blood samples were collected from healthy fertile donors (n\u2009=\u20096). Isolated endometrial cells were cultured using a three-layer gradient system to generate MB-organoids or selected for deriving MB-ESCs. MB-organoids were characterized based on morphological features, including periodic acid\u2013Schiff (PAS) staining for glycogen deposition, scanning electron microscopy (SEM) for pinopode analysis, and immunofluorescence for epithelial (CK8\/18) and stromal (vimentin) markers. ESCs were assessed for decidualization by measuring IGFBP-1 and ZBTB16 expression after hormonal stimulation, with mifepristone used to terminate progesterone signaling.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>MAIN RESULTS AND THE ROLE OF CHANCE<\/jats:title>\n                    <jats:p>MB-organoids demonstrated structural and functional characteristics similar to biopsy-derived endometrial organoids, including glycogen accumulation and pinopode formation, indicative of endometrial receptivity. Immunofluorescence confirmed the presence of both epithelial and stromal populations as well as glycodelin A production. MB-ESCs underwent decidualization in response to hormonal stimulation, with IGFBP-1 and ZBTB16 upregulation, which was suppressed by mifepristone, confirming their functional relevance.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>LIMITATIONS, REASONS FOR CAUTION<\/jats:title>\n                    <jats:p>This in vitro culture system models key endometrial features but lacks the complexity of in vivo conditions. While menstrual blood derived organoids and ESCs respond to hormonal cues, donor variability and the absence of immune and vascular components limit their physiological relevance. Larger studies and more advanced co-culture systems are needed to improve reproducibility and better replicate the dynamic endometrial environment.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>WIDER IMPLICATIONS OF THE FINDINGS<\/jats:title>\n                    <jats:p>Menstrual blood provides a non-invasive, accessible source for generating functional endometrial models. MB-organoids and MB-ESCs offer promising applications in reproductive medicine, including drug screening, disease modeling, and personalized therapies for endometrial disorders.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>STUDY FUNDING\/COMPETING INTEREST(S)<\/jats:title>\n                    <jats:p>This work is supported by the Italian Ministry of University and Research\u2014NextGenerationEU PNRR \u00abTHE\u00bb (Tuscany Health Ecosystem), Spoke 6\u2014Precision Medicine &amp; Personalized Healthcare ECS_00000017, the Estonian Research Council grant no. PRG1076, Swedish Research Council grant no. 2024-02530, Novo Nordisk Foundation grant no. NNF24OC0092384, and Horizon Europe grant NESTOR, grant no. 101120075. The University of Siena Open Access funding partially supported the APC fees. The authors declare no competing interests.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1093\/hropen\/hoaf063","type":"journal-article","created":{"date-parts":[[2025,10,13]],"date-time":"2025-10-13T12:14:32Z","timestamp":1760357672000},"source":"Crossref","is-referenced-by-count":3,"title":["Modeling the window of implantation: insights from endometrial biopsy and menstrual blood-derived organoids and endometrial stromal cells"],"prefix":"10.1093","volume":"2025","author":[{"given":"Francesca Paola","family":"Luongo","sequence":"first","affiliation":[{"name":"Department of Molecular and Developmental Medicine, University of Siena , Siena,","place":["Italy"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Irene","family":"Ortega Ba\u00f1o","sequence":"additional","affiliation":[{"name":"Department of Molecular and Developmental Medicine, University of Siena , Siena,","place":["Italy"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Giuseppe","family":"Belmonte","sequence":"additional","affiliation":[{"name":"Department of Molecular and Developmental Medicine, University of Siena , Siena,","place":["Italy"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mariangela","family":"Gentile","sequence":"additional","affiliation":[{"name":"Department of Life Sciences, University of Siena , Siena,","place":["Italy"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eugenio","family":"Paccagnini","sequence":"additional","affiliation":[{"name":"Department of Life Sciences, University of Siena , Siena,","place":["Italy"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1251-8160","authenticated-orcid":false,"given":"Andres","family":"Salumets","sequence":"additional","affiliation":[{"name":"Division of Obstetrics and Gynaecology, Department of Clinical Science, Intervention and Technology (CLINTEC), Karolinska Institute and Karolinska University Hospital , Stockholm,","place":["Sweden"]},{"name":"Department of Obstetrics and Gynaecology, Institute of Clinical Medicine, University of Tartu , Tartu,","place":["Estonia"]},{"name":"Celvia CC AS (Competence Centre on Health Technologies), Research Institute , Tartu,","place":["Estonia"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2987-7028","authenticated-orcid":false,"given":"Paola","family":"Piomboni","sequence":"additional","affiliation":[{"name":"Department of Molecular and Developmental Medicine, University of Siena , Siena,","place":["Italy"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4946-7336","authenticated-orcid":false,"given":"Alice","family":"Luddi","sequence":"additional","affiliation":[{"name":"Department of Molecular and Developmental Medicine, University of Siena , Siena,","place":["Italy"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2025,10,15]]},"reference":[{"key":"2025111910223166200_hoaf063-B1","doi-asserted-by":"crossref","first-page":"32","DOI":"10.1186\/s13287-015-0013-5","article-title":"Characterization of menstrual stem cells: angiogenic effect, migration and hematopoietic stem cell support in comparison with bone marrow mesenchymal stem cells","volume":"6","author":"Alcayaga-Miranda","year":"2015","journal-title":"Stem Cell Res Ther"},{"key":"2025111910223166200_hoaf063-B2","doi-asserted-by":"crossref","first-page":"836","DOI":"10.1093\/humrep\/16.5.836","article-title":"Endometrial stromal cells regulate epithelial cell growth in vitro: a new co-culture model","volume":"16","author":"Arnold","year":"2001","journal-title":"Hum Reprod"},{"key":"2025111910223166200_hoaf063-B3","doi-asserted-by":"crossref","first-page":"1775","DOI":"10.1242\/dev.148478","article-title":"Development of organoids from mouse and human endometrium showing endometrial epithelium physiology and long-term expandability","volume":"144","author":"Boretto","year":"2017","journal-title":"Development"},{"key":"2025111910223166200_hoaf063-B4","doi-asserted-by":"crossref","first-page":"1041","DOI":"10.1038\/s41556-019-0360-z","article-title":"Patient-derived organoids from endometrial disease capture clinical heterogeneity and are amenable to drug screening","volume":"21","author":"Boretto","year":"2019","journal-title":"Nat Cell Biol"},{"key":"2025111910223166200_hoaf063-B5","doi-asserted-by":"crossref","first-page":"3894","DOI":"10.1038\/srep03894","article-title":"Uterine selection of human embryos at implantation","volume":"4","author":"Brosens","year":"2014","journal-title":"Sci Rep"},{"key":"2025111910223166200_hoaf063-B6","doi-asserted-by":"crossref","first-page":"651","DOI":"10.1038\/s42003-021-02194-y","article-title":"Menstrual flow as a non-invasive source of endometrial organoids","volume":"4","author":"Cindrova-Davies","year":"2021","journal-title":"Commun Biol"},{"key":"2025111910223166200_hoaf063-B7","doi-asserted-by":"crossref","first-page":"1236","DOI":"10.3390\/cells13151236","article-title":"Unraveling the dynamics of estrogen and progesterone signaling in the endometrium: an overview","volume":"13","author":"Dias Da Silva","year":"2024","journal-title":"Cells"},{"key":"2025111910223166200_hoaf063-B8","doi-asserted-by":"crossref","first-page":"23132","DOI":"10.1073\/pnas.1915389116","article-title":"Self-renewing endometrial epithelial organoids of the human uterus","volume":"116","author":"Fitzgerald","year":"2019","journal-title":"Proc Natl Acad Sci U S A"},{"key":"2025111910223166200_hoaf063-B9","doi-asserted-by":"crossref","first-page":"579","DOI":"10.1177\/1933719117718276","article-title":"Dysregulation of GdA expression in endometrium of women with endometriosis: implication for endometrial receptivity","volume":"25","author":"Focarelli","year":"2018","journal-title":"Reprod Sci"},{"key":"2025111910223166200_hoaf063-B10","doi-asserted-by":"crossref","first-page":"862","DOI":"10.1093\/humrep\/dep468","article-title":"Invasiveness of human endometrial stromal cells is promoted by decidualization and by trophoblast-derived signals","volume":"25","author":"Gellersen","year":"2010","journal-title":"Hum Reprod"},{"key":"2025111910223166200_hoaf063-B11","doi-asserted-by":"crossref","first-page":"101656","DOI":"10.1016\/j.tice.2021.101656","article-title":"Main actors behind the endometrial receptivity and successful implantation","volume":"73","author":"Governini","year":"2021","journal-title":"Tissue Cell"},{"key":"2025111910223166200_hoaf063-B12","doi-asserted-by":"crossref","first-page":"408","DOI":"10.1177\/1933719113497284","article-title":"Mifepristone-exposured human endometrial endothelial cells in vitro","volume":"21","author":"Helmestam","year":"2014","journal-title":"Reprod Sci"},{"key":"2025111910223166200_hoaf063-B13","doi-asserted-by":"crossref","first-page":"793","DOI":"10.1093\/oxfordjournals.humrep.a138144","article-title":"The production of placental protein 14 and interleukin 6 by human endometrial cells in culture","volume":"8","author":"Laird","year":"1993","journal-title":"Hum Reprod"},{"key":"2025111910223166200_hoaf063-B14","doi-asserted-by":"crossref","first-page":"3031","DOI":"10.1093\/humrep\/dem297","article-title":"Mifepristone, but not levonorgestrel, inhibits human blastocyst attachment to an in vitro endometrial three-dimensional cell culture model","volume":"22","author":"Lalitkumar","year":"2007","journal-title":"Hum Reprod"},{"key":"2025111910223166200_hoaf063-B15","doi-asserted-by":"crossref","first-page":"37","DOI":"10.1055\/s-2001-13909","article-title":"On the role of human chorionic gonadotropin (hCG) in the embryo-endometrial microenvironment: implications for differentiation and implantation","volume":"19","author":"Licht","year":"2001","journal-title":"Semin Reprod Med"},{"key":"2025111910223166200_hoaf063-B16","doi-asserted-by":"crossref","first-page":"37","DOI":"10.1038\/s42003-020-0763-1","article-title":"Recurrent pregnancy loss is associated with a pro-senescent decidual response during the peri-implantation window","volume":"3","author":"Lucas","year":"2020","journal-title":"Commun Biol"},{"key":"2025111910223166200_hoaf063-B17","doi-asserted-by":"crossref","first-page":"1054","DOI":"10.1016\/j.fertnstert.2020.10.058","article-title":"Emerging role of embryo secretome in the paracrine communication at the implantation site: a proof of concept","volume":"115","author":"Luddi","year":"2021","journal-title":"Fertil Steril"},{"key":"2025111910223166200_hoaf063-B18","doi-asserted-by":"crossref","first-page":"1121","DOI":"10.3390\/cells9051121","article-title":"Organoids of human endometrium: a powerful in vitro model for the endometrium-embryo cross-talk at the implantation site","volume":"9","author":"Luddi","year":"2020","journal-title":"Cells"},{"key":"2025111910223166200_hoaf063-B19","doi-asserted-by":"crossref","first-page":"2349","DOI":"10.3390\/cells12192349","article-title":"Exposure to cumulus cell secretome improves sperm function: new perspectives for sperm selection in vitro","volume":"12","author":"Luongo","year":"2023","journal-title":"Cells"},{"key":"2025111910223166200_hoaf063-B20","doi-asserted-by":"crossref","first-page":"45","DOI":"10.1080\/09513590.2019.1653562","article-title":"Ultrastructural markers of tissue endometrial receptivity in patients with recurrent implantation failure","volume":"35","author":"Melkozerova","year":"2019","journal-title":"Gynecol Endocrinol"},{"key":"2025111910223166200_hoaf063-B21","first-page":"230","article-title":"Biological mechanisms underlying the clinical effects of mifepristone (RU 486) on the endometrium","volume":"4","author":"Papp","year":"2000","journal-title":"Early Pregnancy (Cherry Hill)"},{"key":"2025111910223166200_hoaf063-B22","first-page":"849","article-title":"3D model of the maternal-fetal interface: challenges, recent advances and beyond","volume":"38","author":"Passaponti","year":"2023","journal-title":"Histol Histopathol"},{"key":"2025111910223166200_hoaf063-B23","doi-asserted-by":"crossref","first-page":"303","DOI":"10.3727\/096368908784153922","article-title":"Multipotent menstrual blood stromal stem cells: isolation, characterization, and differentiation","volume":"17","author":"Patel","year":"2008","journal-title":"Cell Transplant"},{"key":"2025111910223166200_hoaf063-B24","doi-asserted-by":"crossref","first-page":"2633","DOI":"10.1093\/humrep\/deh450","article-title":"Human chorionic gonadotropin and growth factors at the embryonic\u2013endometrial interface control leukemia inhibitory factor (LIF) and interleukin 6 (IL-6) secretion by human endometrial epithelium","volume":"19","author":"Perrier d\u2019Hauterive","year":"2004","journal-title":"Hum Reprod"},{"key":"2025111910223166200_hoaf063-B25","doi-asserted-by":"crossref","first-page":"e40","DOI":"10.4142\/jvs.24004","article-title":"Current strategies using 3D organoids to establish in vitro maternal-embryonic interaction","volume":"25","author":"Saadeldin","year":"2024","journal-title":"J Vet Sci"},{"key":"2025111910223166200_hoaf063-B26","doi-asserted-by":"crossref","first-page":"401","DOI":"10.1210\/er.2001-0026","article-title":"Glycodelin: a major lipocalin protein of the reproductive axis with diverse actions in cell recognition and differentiation","volume":"23","author":"Sepp\u00e4l\u00e4","year":"2002","journal-title":"Endocr Rev"},{"key":"2025111910223166200_hoaf063-B27","doi-asserted-by":"crossref","first-page":"eadi4819","DOI":"10.1126\/sciadv.adi4819","article-title":"Modeling embryo-endometrial interface recapitulating human embryo implantation","volume":"10","author":"Shibata","year":"2024","journal-title":"Sci Adv"},{"key":"2025111910223166200_hoaf063-B28","doi-asserted-by":"crossref","first-page":"568","DOI":"10.1038\/ncb3516","article-title":"Long-term, hormone-responsive organoid cultures of human endometrium in a chemically defined medium","volume":"19","author":"Turco","year":"2017","journal-title":"Nat Cell Biol"},{"key":"2025111910223166200_hoaf063-B29","doi-asserted-by":"crossref","first-page":"108522","DOI":"10.1016\/j.reprotox.2023.108522","article-title":"Long-term maintenance of human endometrial epithelial organoids and their stem cell properties","volume":"123","author":"Zhang","year":"2024","journal-title":"Reprod Toxicol"},{"key":"2025111910223166200_hoaf063-B30","article-title":"Human receptive endometrial organoid for deciphering the implantation window","volume":"12","author":"Zhang","year":"2023","journal-title":"eLife"}],"container-title":["Human Reproduction Open"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/academic.oup.com\/hropen\/advance-article-pdf\/doi\/10.1093\/hropen\/hoaf063\/64711660\/hoaf063.pdf","content-type":"application\/pdf","content-version":"am","intended-application":"syndication"},{"URL":"https:\/\/academic.oup.com\/hropen\/article-pdf\/2025\/4\/hoaf063\/64711660\/hoaf063.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/academic.oup.com\/hropen\/article-pdf\/2025\/4\/hoaf063\/64711660\/hoaf063.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,11,19]],"date-time":"2025-11-19T15:22:42Z","timestamp":1763565762000},"score":1,"resource":{"primary":{"URL":"https:\/\/academic.oup.com\/hropen\/article\/doi\/10.1093\/hropen\/hoaf063\/8286935"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025]]},"references-count":30,"journal-issue":{"issue":"4","published-print":{"date-parts":[[2025,9,22]]}},"URL":"https:\/\/doi.org\/10.1093\/hropen\/hoaf063","relation":{},"ISSN":["2399-3529"],"issn-type":[{"value":"2399-3529","type":"electronic"}],"subject":[],"published-other":{"date-parts":[[2025]]},"published":{"date-parts":[[2025]]},"article-number":"hoaf063"}}