A protocol for organoids from the gynecomastia patients

BackgroundGynecomastia, 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.MethodsIn this study, we established...

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Main Authors: Fangjian Shang, Zihao Li, Ji Feng, Qi Wang, Mengyang An, Zengren Zhao, Bo Liu
Format: Article
Language:English
Published: Frontiers Media S.A. 2025-08-01
Series:Frontiers in Bioengineering and Biotechnology
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Online Access:https://www.frontiersin.org/articles/10.3389/fbioe.2025.1593368/full
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author Fangjian Shang
Zihao Li
Ji Feng
Qi Wang
Mengyang An
Zengren Zhao
Bo Liu
author_facet Fangjian Shang
Zihao Li
Ji Feng
Qi Wang
Mengyang An
Zengren Zhao
Bo Liu
author_sort Fangjian Shang
collection DOAJ
description BackgroundGynecomastia, 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.MethodsIn 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.ResultsThe organoids recapitulated key histological and molecular features of their corresponding source tissues, including epithelial architecture and expression of CK14, CK18, Ki67, and ERα. Marker expression was generally consistent between organoids and tissues at both the protein and transcriptional levels. Notably, ERα protein levels were reduced in organoids, while ESR1 mRNA expression remained stable, suggesting post-transcriptional regulation related to culture conditions.ConclusionOur 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.
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spelling doaj-art-e2d275bfefcb4c7bbcbf831c8749753d2025-08-20T04:00:54ZengFrontiers Media S.A.Frontiers in Bioengineering and Biotechnology2296-41852025-08-011310.3389/fbioe.2025.15933681593368A protocol for organoids from the gynecomastia patientsFangjian ShangZihao LiJi FengQi WangMengyang AnZengren ZhaoBo LiuBackgroundGynecomastia, 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.MethodsIn 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.ResultsThe organoids recapitulated key histological and molecular features of their corresponding source tissues, including epithelial architecture and expression of CK14, CK18, Ki67, and ERα. Marker expression was generally consistent between organoids and tissues at both the protein and transcriptional levels. Notably, ERα protein levels were reduced in organoids, while ESR1 mRNA expression remained stable, suggesting post-transcriptional regulation related to culture conditions.ConclusionOur 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.https://www.frontiersin.org/articles/10.3389/fbioe.2025.1593368/fullgynecomastiaorganoid cultureimmunohistochemistry and immunofluorescence analysisqPCR validation3D disease modelinghormone receptor signaling
spellingShingle Fangjian Shang
Zihao Li
Ji Feng
Qi Wang
Mengyang An
Zengren Zhao
Bo Liu
A protocol for organoids from the gynecomastia patients
Frontiers in Bioengineering and Biotechnology
gynecomastia
organoid culture
immunohistochemistry and immunofluorescence analysis
qPCR validation
3D disease modeling
hormone receptor signaling
title A protocol for organoids from the gynecomastia patients
title_full A protocol for organoids from the gynecomastia patients
title_fullStr A protocol for organoids from the gynecomastia patients
title_full_unstemmed A protocol for organoids from the gynecomastia patients
title_short A protocol for organoids from the gynecomastia patients
title_sort protocol for organoids from the gynecomastia patients
topic gynecomastia
organoid culture
immunohistochemistry and immunofluorescence analysis
qPCR validation
3D disease modeling
hormone receptor signaling
url https://www.frontiersin.org/articles/10.3389/fbioe.2025.1593368/full
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