METHODS article
Front. Bioeng. Biotechnol.
Sec. Organoids and Organ-On-A-Chip
Volume 13 - 2025 | doi: 10.3389/fbioe.2025.1593368
A Protocol for Organoids from the Gynecomastia Patients
Provisionally accepted
- The First hospital of Hebei Medical University, Shijiazhuang, China
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Background: Gynecomastia, characterized by benign proliferation of male breast glandular tissue, is a prevalent condition with complex etiologies. However, the absence of effective in vitro models has hindered mechanistic investigations and therapeutic development. Methods: 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&E) staining, immunohistochemistry (IHC), immunofluorescence (IF), and quantitative PCR (qPCR), targeting a panel of lineage-specific and proliferative markers.The organoids recapitulated key histological and molecular features of their corresponding source tissues, including epithelial architecture and expression of CK14, CK18, Ki67, and ER α . Marker expression was generally consistent between organoids and tissues at both the protein and transcriptional levels. Notably, ER α protein levels were reduced in organoids, while ESR1 mRNA expression remained stable, suggesting post-transcriptional regulation related to culture conditions. Conclusion: 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.
Keywords: Gynecomastia, organoid culture, Immunohistochemistry and Immunofluorescence Analysis, qPCR validation, 3D disease modeling, Hormone receptor signaling
Received: 19 Mar 2025; Accepted: 31 Jul 2025.
Copyright: © 2025 Shang, Li, Feng, Wang, An, Zhao and Liu. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence:
Zengren Zhao, The First hospital of Hebei Medical University, Shijiazhuang, China
Bo Liu, The First hospital of Hebei Medical University, Shijiazhuang, China
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