{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,12,4]],"date-time":"2025-12-04T18:51:18Z","timestamp":1764874278456,"version":"3.46.0"},"reference-count":25,"publisher":"Public Library of Science (PLoS)","issue":"12","license":[{"start":{"date-parts":[[2025,12,4]],"date-time":"2025-12-04T00:00:00Z","timestamp":1764806400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"the Office of the Director, National Institutes of Health","award":["K01OD032455"],"award-info":[{"award-number":["K01OD032455"]}]}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS One"],"abstract":"<jats:p>\n                    Oviductal (fallopian tube) cell cultures are widely used to improve understanding of oviductal pathophysiology and to improve sperm and embryo quality via co-culture. One type of oviductal cell culture is organoids, a three-dimensional cell culture that can mimic the oviduct\n                    <jats:italic>in vitro<\/jats:italic>\n                    long-term (months). Because organoids maintain their structure and function similar to\n                    <jats:italic>in vivo<\/jats:italic>\n                    , their secretions tend to be more similar to\n                    <jats:italic>in vivo<\/jats:italic>\n                    cellular products. One of these organoid secretions includes extracellular vesicles (EVs), which are bioactive nanoparticles secreted by cells for intercellular communication. Here we report a protocol for isolation of mouse oviducts, establishment and long-term culture of mouse oviductal organoids, and isolation and characterization of the EVs secreted by mouse oviductal organoids. This lab protocol can be used for research of oviductal physiology and pathology and improvement of assisted reproductive technologies.\n                  <\/jats:p>","DOI":"10.1371\/journal.pone.0337587","type":"journal-article","created":{"date-parts":[[2025,12,4]],"date-time":"2025-12-04T18:40:09Z","timestamp":1764873609000},"page":"e0337587","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":0,"title":["Establishment and culture of mouse oviductal organoids and isolation and characterization of their secreted extracellular vesicles"],"prefix":"10.1371","volume":"20","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-1592-1307","authenticated-orcid":true,"given":"Riley E.","family":"Thompson-Brandhagen","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mindy A.","family":"Meyers","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2404-2603","authenticated-orcid":true,"given":"Richard B.","family":"McCosh","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Fiona K.","family":"Hollinshead","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"340","published-online":{"date-parts":[[2025,12,4]]},"reference":[{"issue":"6","key":"pone.0337587.ref001","doi-asserted-by":"crossref","first-page":"3186","DOI":"10.3390\/ijms23063186","article-title":"The Roles of Extracellular Vesicles and Organoid Models in Female Reproductive Physiology","volume":"23","author":"RE Thompson","year":"2022","journal-title":"Int J Mol 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chemically defined medium","volume":"19","author":"MY Turco","year":"2017","journal-title":"Nat Cell Biol"},{"issue":"7735","key":"pone.0337587.ref006","doi-asserted-by":"crossref","first-page":"263","DOI":"10.1038\/s41586-018-0753-3","article-title":"Trophoblast organoids as a model for maternal-fetal interactions during human placentation","volume":"564","author":"MY Turco","year":"2018","journal-title":"Nature"},{"issue":"10","key":"pone.0337587.ref007","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":"M Boretto","year":"2017","journal-title":"Development"},{"key":"pone.0337587.ref008","doi-asserted-by":"crossref","first-page":"51","DOI":"10.1016\/j.scr.2018.08.021","article-title":"Long-term organoid culture reveals enrichment of organoid-forming epithelial cells in the fimbrial portion of mouse fallopian tube","volume":"32","author":"Y Xie","year":"2018","journal-title":"Stem Cell Res"},{"issue":"3","key":"pone.0337587.ref009","doi-asserted-by":"crossref","first-page":"503","DOI":"10.1093\/humrep\/deae258","article-title":"Human fallopian tube organoids provide a favourable environment for sperm motility","volume":"40","author":"N Gatimel","year":"2025","journal-title":"Human Reproduction"},{"issue":"1","key":"pone.0337587.ref010","doi-asserted-by":"crossref","first-page":"5367","DOI":"10.1038\/s41467-019-13116-2","article-title":"Both fallopian tube and ovarian surface epithelium are cells-of-origin for high-grade serous ovarian carcinoma","volume":"10","author":"S Zhang","year":"2019","journal-title":"Nat Commun"},{"issue":"1","key":"pone.0337587.ref011","doi-asserted-by":"crossref","first-page":"646","DOI":"10.1186\/s12864-023-09746-y","article-title":"Bovine oviductal organoids: a multi-omics approach to capture the cellular and extracellular molecular response of the oviduct to heat stress","volume":"24","author":"NG Menjivar","year":"2023","journal-title":"BMC Genomics"},{"issue":"1","key":"pone.0337587.ref012","doi-asserted-by":"crossref","first-page":"1535750","DOI":"10.1080\/20013078.2018.1535750","article-title":"Minimal information for studies of extracellular vesicles 2018 (MISEV2018): a position statement of the International Society for Extracellular Vesicles and update of the MISEV2014 guidelines","volume":"7","author":"C Th\u00e9ry","year":"2018","journal-title":"J Extracell Vesicles"},{"issue":"11","key":"pone.0337587.ref013","doi-asserted-by":"crossref","first-page":"276","DOI":"10.3390\/metabo9110276","article-title":"Metabolomics Applied to the Study of Extracellular Vesicles","volume":"9","author":"C Williams","year":"2019","journal-title":"Metabolites"},{"issue":"1","key":"pone.0337587.ref014","doi-asserted-by":"crossref","first-page":"501","DOI":"10.1038\/s41598-019-57060-z","article-title":"Amniotic microvesicles impact 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