{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,24]],"date-time":"2026-10-24T00:27:58Z","timestamp":1792801678001,"version":"4.5.0"},"reference-count":11,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2025,8,13]],"date-time":"2025-08-13T00:00:00Z","timestamp":1755043200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Bioeng. Biotechnol."],"abstract":"<jats:sec><jats:title>Background<\/jats:title><jats:p>Gynecomastia, characterized by benign proliferation of male breast glandular tissue, is a prevalent condition with complex etiologies. However, the absence of effective <jats:italic>in vitro<\/jats:italic> models has hindered mechanistic investigations and therapeutic development.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>In this study, we established and characterized organoids derived from the breast tissues of six male gynecomastia patients, including physiological, idiopathic, and hormone-related subtypes. Organoid fidelity was evaluated using hematoxylin and eosin (H&amp;amp;E) staining, immunohistochemistry (IHC), immunofluorescence (IF), and quantitative PCR (qPCR), targeting a panel of lineage-specific and proliferative markers.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>The organoids recapitulated key histological and molecular features of their corresponding source tissues, including epithelial architecture and expression of CK14, CK18, Ki67, and ER\u03b1. Marker expression was generally consistent between organoids and tissues at both the protein and transcriptional levels. Notably, ER\u03b1 protein levels were reduced in organoids, while ESR1 mRNA expression remained stable, suggesting post-transcriptional regulation related to culture conditions.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>Our study presents a practical and reproducible protocol for generating gynecomastia-derived organoids and highlights their utility as a disease-relevant platform for future research in male breast pathology and hormone-related mechanisms.<\/jats:p><\/jats:sec>","DOI":"10.3389\/fbioe.2025.1593368","type":"journal-article","created":{"date-parts":[[2025,8,13]],"date-time":"2025-08-13T10:40:17Z","timestamp":1755081617000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":1,"title":["A protocol for organoids from the gynecomastia patients"],"prefix":"10.3389","volume":"13","author":[{"given":"Fangjian","family":"Shang","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zihao","family":"Li","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ji","family":"Feng","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qi","family":"Wang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mengyang","family":"An","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zengren","family":"Zhao","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bo","family":"Liu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2025,8,13]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"150","DOI":"10.4103\/2230-8210.129104","article-title":"Gynecomastia: clinical evaluation and management","volume":"18","author":"Cuhaci","year":"2014","journal-title":"Indian J. Endocrinol. Metab."},{"key":"B2","doi-asserted-by":"publisher","first-page":"1756","DOI":"10.1038\/s41596-019-0160-8","article-title":"High-resolution 3D imaging of fixed and cleared organoids","volume":"14","author":"Dekkers","year":"2019","journal-title":"Nat. Protoc."},{"key":"B3","doi-asserted-by":"publisher","first-page":"471","DOI":"10.1634\/theoncologist.10-7-471","article-title":"A review of the diagnosis and management of Male breast cancer","volume":"10","author":"Giordano","year":"2005","journal-title":"Oncologist"},{"key":"B4","doi-asserted-by":"publisher","first-page":"716","DOI":"10.1016\/S0025-6196(11)60671-X","article-title":"Gynecomastia: pathophysiology, evaluation, and management","volume":"80","author":"Johnson","year":"2009","journal-title":"Am. Fam. 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Adolesc."},{"key":"B8","doi-asserted-by":"publisher","first-page":"373","DOI":"10.1016\/j.cell.2017.11.010","article-title":"A living biobank of breast cancer organoids captures disease heterogeneity","volume":"172","author":"Sachs","year":"2018","journal-title":"Cell."},{"key":"B9","doi-asserted-by":"publisher","first-page":"159","DOI":"10.1016\/j.surg.2023.04.022","article-title":"The combination of endoscopic subcutaneous mastectomy and liposuction (Liu and Shang's 2-hole 7-step method) as the treatment of gynecomastia","volume":"174","author":"Shang","year":"2023","journal-title":"Surgery"},{"key":"B10","doi-asserted-by":"publisher","first-page":"98","DOI":"10.1186\/1471-2121-11-98","article-title":"Ligands specify estrogen receptor alpha nuclear localization and degradation","volume":"11","author":"S\u00f6derqvist","year":"2010","journal-title":"BMC Cell. Biol."},{"key":"B11","doi-asserted-by":"publisher","first-page":"94","DOI":"10.1038\/s43586-022-00174-y","article-title":"Organoids","volume":"2","author":"Zhao","year":"2022","journal-title":"Nat. Rev. Methods Prim."}],"container-title":["Frontiers in Bioengineering and Biotechnology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fbioe.2025.1593368\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,8,13]],"date-time":"2025-08-13T10:40:18Z","timestamp":1755081618000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fbioe.2025.1593368\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,8,13]]},"references-count":11,"alternative-id":["10.3389\/fbioe.2025.1593368"],"URL":"https:\/\/doi.org\/10.3389\/fbioe.2025.1593368","relation":{},"ISSN":["2296-4185"],"issn-type":[{"value":"2296-4185","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,8,13]]},"article-number":"1593368"}}