{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,12]],"date-time":"2026-10-12T16:49:20Z","timestamp":1791823760634,"version":"4.3.4"},"reference-count":38,"publisher":"Wiley","issue":"6","license":[{"start":{"date-parts":[[2023,11,14]],"date-time":"2023-11-14T00:00:00Z","timestamp":1699920000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["American J Rep Immunol"],"published-print":{"date-parts":[[2023,12]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>The vaginal microbiome includes diverse microbiota dominated by <jats:italic>Lactobacillus [L.] spp<\/jats:italic>. that protect against infections, modulate inflammation, and regulate vaginal homeostasis. Because it is challenging to incorporate vaginal microbiota into in vitro models, including organ\u2010on\u2010a\u2010chip systems, we assessed microbial metabolites as reliable proxies in addition to traditional vaginal epithelial cultures (VECs). Human immortalized VECs cultured on transwells with an air\u2010liquid interface generated stratified cell layers colonized by transplanted healthy microbiomes (<jats:italic>L. jensenii\u2010<\/jats:italic> or <jats:italic>L. crispatus<\/jats:italic>\u2010dominant) or a community representing bacterial vaginosis (BV). After 48\u2010h, a qPCR array confirmed the expected donor community profiles. Pooled apical and basal supernatants were subjected to metabolomic analysis (untargeted mass spectrometry) followed by ingenuity pathways analysis (IPA). To determine the bacterial metabolites\u2019 ability to recreate the vaginal microenvironment in vitro, pooled bacteria\u2010free metabolites were added to traditional VEC cultures. Cell morphology, viability, and cytokine production were assessed. IPA analysis of metabolites from colonized samples contained fatty acids, nucleic acids, and sugar acids that were associated with signaling networks that contribute to secondary metabolism, anti\u2010fungal, and anti\u2010inflammatory functions indicative of a healthy vaginal microbiome compared to sterile VEC transwell metabolites. Pooled metabolites did not affect cell morphology or induce cell death (\u223c5.5%) of VEC cultures (<jats:italic>n<\/jats:italic>\u00a0=\u00a03) after 72\u2010h. However, metabolites created an anti\u2010inflammatory milieu by increasing IL\u201010 production (<jats:italic>p<\/jats:italic>\u00a0=\u00a0.06, <jats:italic>T<\/jats:italic>\u2010test) and significantly suppressing pro\u2010inflammatory IL\u20106 (<jats:italic>p<\/jats:italic>\u00a0=\u00a0.0001), IL\u20108 (<jats:italic>p<\/jats:italic>\u00a0=\u00a0.009), and TNF\u03b1 (<jats:italic>p<\/jats:italic>\u00a0=\u00a0.0007) compared to na\u00efve VEC cultures. BV VEC conditioned\u2010medium did not affect cell morphology nor viability; however, it induced a pro\u2010inflammatory environment by elevating levels of IL\u20106 (<jats:italic>p<\/jats:italic>\u00a0=\u00a0.023), IL\u20108 (<jats:italic>p<\/jats:italic>\u00a0=\u00a0.031), and TNF\u03b1 (<jats:italic>p<\/jats:italic>\u00a0=\u00a0.021) when compared to <jats:italic>L<\/jats:italic>.\u2010dominate microbiome\u2010conditioned medium. VEC transwells provide a suitable ex vivo system to support the production of bacterial metabolites consistent with the vaginal milieu allowing subsequent in vitro studies with enhanced accuracy and utility.<\/jats:p>","DOI":"10.1111\/aji.13797","type":"journal-article","created":{"date-parts":[[2023,11,15]],"date-time":"2023-11-15T20:22:51Z","timestamp":1700079771000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":8,"title":["Validation of vaginal microbiome proxies for\u00a0in vitro\u00a0experiments that biomimic Lactobacillus\u2010dominant vaginal cultures"],"prefix":"10.1111","volume":"90","author":[{"given":"Abir","family":"Zahra","sequence":"first","affiliation":[{"name":"School of Medicine The University of Texas Medical Branch at Galveston  Galveston Texas USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9213-6105","authenticated-orcid":false,"given":"Ramkumar","family":"Menon","sequence":"additional","affiliation":[{"name":"Division of Basic Science and Translational Research Department of Obstetrics and Gynecology The University of Texas Medical Branch at Galveston  Galveston Texas USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3868-2935","authenticated-orcid":false,"given":"Giovana Fernanda Cosi","family":"Bento","sequence":"additional","affiliation":[{"name":"Division of Basic Science and Translational Research Department of Obstetrics and Gynecology The University of Texas Medical Branch at Galveston  Galveston Texas USA"},{"name":"Department of Pathology Botucatu Medical School S\u00e3o Paulo State University  S\u00e3o Paulo Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jessica","family":"Selim","sequence":"additional","affiliation":[{"name":"School of Medicine The University of Texas Medical Branch at Galveston  Galveston Texas USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8234-1815","authenticated-orcid":false,"given":"Brandie D.","family":"Taylor","sequence":"additional","affiliation":[{"name":"Division of Basic Science and Translational Research Department of Obstetrics and Gynecology The University of Texas Medical Branch at Galveston  Galveston Texas USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kathleen L.","family":"Vincent","sequence":"additional","affiliation":[{"name":"Division of Basic Science and Translational Research Department of Obstetrics and Gynecology The University of Texas Medical Branch at Galveston  Galveston Texas USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Richard B.","family":"Pyles","sequence":"additional","affiliation":[{"name":"Department of Pediatrics The University of Texas Medical Branch  Galveston Texas USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8392-2833","authenticated-orcid":false,"given":"Lauren S.","family":"Richardson","sequence":"additional","affiliation":[{"name":"Division of Basic Science and Translational Research Department of Obstetrics and Gynecology The University of Texas Medical Branch at Galveston  Galveston Texas 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