Comparative analysis of organoid, air-liquid interface, and direct infection models for studying pathogen–host interactions in endometrial tissue

Abstract The endometrium is a critical component of female reproductive health. Endometritis can significantly affect women’s health, leading to complications such as infertility, pregnancy failure, and intrauterine adhesions. Therefore, establishing a reliable and effective model of endometritis is...

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Main Authors: Xin Zhang, Linyuan Fan, Li Zhang, Zhaohui Liu
Format: Article
Language:English
Published: Nature Portfolio 2025-03-01
Series:Scientific Reports
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Online Access:https://doi.org/10.1038/s41598-025-93374-x
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author Xin Zhang
Linyuan Fan
Li Zhang
Zhaohui Liu
author_facet Xin Zhang
Linyuan Fan
Li Zhang
Zhaohui Liu
author_sort Xin Zhang
collection DOAJ
description Abstract The endometrium is a critical component of female reproductive health. Endometritis can significantly affect women’s health, leading to complications such as infertility, pregnancy failure, and intrauterine adhesions. Therefore, establishing a reliable and effective model of endometritis is essential for advancing research in female reproductive health. The air-liquid interface culture models epithelial exposure to both air and medium, supporting the study of apical-basal polarity and immune responses, but lacks the three-dimensional structure of organoids. Microinjection delivers bacteria directly into the organoid lumen, facilitating the study of bacterial invasion and replication, though it requires advanced technical skills and may not fully replicate natural infections. The direct infection of organoids in suspension culture offers a more realistic model of ascending infections by exposing the basal epithelial surfaces to bacteria, more closely mimicking in vivo conditions. Among the models tested, the direct infection method most accurately mirrors the progression of E. coli infection, showing its advantage in studying bacterial adhesion, replication, and epithelial barrier disruption. Our comparative analysis of microinjection, direct infection, and the ALI model highlighted distinct advantages and challenges associated with each. Microinjection offers precise delivery but is hindered by technical complexity and equipment demands. The ALI model, despite its efficacy, requires extended culture times and limits direct visualization of cell development. Conversely, the direct infection model, which involves the simple removal of Matrigel, proves to be user-friendly, cost-effective, and permits continuous observation of cell behavior. Nonetheless, the direct infection model still presents certain limitations that warrant further optimization.
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spelling doaj-art-263f575c87bf4e5da88f7b742b025f2a2025-08-20T03:01:23ZengNature PortfolioScientific Reports2045-23222025-03-0115111110.1038/s41598-025-93374-xComparative analysis of organoid, air-liquid interface, and direct infection models for studying pathogen–host interactions in endometrial tissueXin Zhang0Linyuan Fan1Li Zhang2Zhaohui Liu3Department of Gynecology, Beijing Obstetrics and Gynecology Hospital, Capital Medical University, Beijing Maternal and Child Health Care HospitalDepartment of Gynecology, Beijing Obstetrics and Gynecology Hospital, Capital Medical University, Beijing Maternal and Child Health Care HospitalDepartment of Gynecology, Beijing Obstetrics and Gynecology Hospital, Capital Medical University, Beijing Maternal and Child Health Care HospitalDepartment of Gynecology, Beijing Obstetrics and Gynecology Hospital, Capital Medical University, Beijing Maternal and Child Health Care HospitalAbstract The endometrium is a critical component of female reproductive health. Endometritis can significantly affect women’s health, leading to complications such as infertility, pregnancy failure, and intrauterine adhesions. Therefore, establishing a reliable and effective model of endometritis is essential for advancing research in female reproductive health. The air-liquid interface culture models epithelial exposure to both air and medium, supporting the study of apical-basal polarity and immune responses, but lacks the three-dimensional structure of organoids. Microinjection delivers bacteria directly into the organoid lumen, facilitating the study of bacterial invasion and replication, though it requires advanced technical skills and may not fully replicate natural infections. The direct infection of organoids in suspension culture offers a more realistic model of ascending infections by exposing the basal epithelial surfaces to bacteria, more closely mimicking in vivo conditions. Among the models tested, the direct infection method most accurately mirrors the progression of E. coli infection, showing its advantage in studying bacterial adhesion, replication, and epithelial barrier disruption. Our comparative analysis of microinjection, direct infection, and the ALI model highlighted distinct advantages and challenges associated with each. Microinjection offers precise delivery but is hindered by technical complexity and equipment demands. The ALI model, despite its efficacy, requires extended culture times and limits direct visualization of cell development. Conversely, the direct infection model, which involves the simple removal of Matrigel, proves to be user-friendly, cost-effective, and permits continuous observation of cell behavior. Nonetheless, the direct infection model still presents certain limitations that warrant further optimization.https://doi.org/10.1038/s41598-025-93374-xInfection modelEndometrial organoidsAir-liquid interfaceTranswell
spellingShingle Xin Zhang
Linyuan Fan
Li Zhang
Zhaohui Liu
Comparative analysis of organoid, air-liquid interface, and direct infection models for studying pathogen–host interactions in endometrial tissue
Scientific Reports
Infection model
Endometrial organoids
Air-liquid interface
Transwell
title Comparative analysis of organoid, air-liquid interface, and direct infection models for studying pathogen–host interactions in endometrial tissue
title_full Comparative analysis of organoid, air-liquid interface, and direct infection models for studying pathogen–host interactions in endometrial tissue
title_fullStr Comparative analysis of organoid, air-liquid interface, and direct infection models for studying pathogen–host interactions in endometrial tissue
title_full_unstemmed Comparative analysis of organoid, air-liquid interface, and direct infection models for studying pathogen–host interactions in endometrial tissue
title_short Comparative analysis of organoid, air-liquid interface, and direct infection models for studying pathogen–host interactions in endometrial tissue
title_sort comparative analysis of organoid air liquid interface and direct infection models for studying pathogen host interactions in endometrial tissue
topic Infection model
Endometrial organoids
Air-liquid interface
Transwell
url https://doi.org/10.1038/s41598-025-93374-x
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AT linyuanfan comparativeanalysisoforganoidairliquidinterfaceanddirectinfectionmodelsforstudyingpathogenhostinteractionsinendometrialtissue
AT lizhang comparativeanalysisoforganoidairliquidinterfaceanddirectinfectionmodelsforstudyingpathogenhostinteractionsinendometrialtissue
AT zhaohuiliu comparativeanalysisoforganoidairliquidinterfaceanddirectinfectionmodelsforstudyingpathogenhostinteractionsinendometrialtissue