{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,9]],"date-time":"2026-09-09T17:44:50Z","timestamp":1788975890907,"version":"build-2803163510"},"reference-count":53,"publisher":"Wiley","issue":"5","license":[{"start":{"date-parts":[[2023,8,28]],"date-time":"2023-08-28T00:00:00Z","timestamp":1693180800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"funder":[{"DOI":"10.13039\/100008980","name":"Medical College of Wisconsin","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100008980","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000055","name":"National Institute on Deafness and Other Communication Disorders","doi-asserted-by":"publisher","award":["5R21DC015634\u201002"],"award-info":[{"award-number":["5R21DC015634\u201002"]}],"id":[{"id":"10.13039\/100000055","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Laryngoscope Investig Oto"],"published-print":{"date-parts":[[2023,10]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:title>Objective<\/jats:title><jats:p>Otitis media (OM) is among the most frequently diagnosed pediatric diseases in the US. Despite the significant public health burden of OM and the contribution research in culture models has made to understanding its pathobiology, a singular immortalized human middle ear epithelial (MEE) cell line exists (HMEEC\u20101, adult\u2010derived). We previously developed MEE cultures from pediatric patients with non\u2010inflamed MEE (PCI), recurrent OM (ROM), or OM with effusion (OME) and demonstrated differences in their baseline inflammatory cytokine expression and response to stimulation with an OM\u2010relevant pathogen lysate and cytokines. Herein, we sought to immortalize these cultures and assess retention of their phenotypes.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>MEE cultures were immortalized via lentivirus encoding temperature\u2010sensitive SV40 T antigen. Immortalized MEE lines and HMEEC\u20101 grown in monolayer were stimulated with non\u2010typeable <jats:italic>Haemophilus influenzae<\/jats:italic> (NTHi) lysate. Gene expression (<jats:italic>TNFA<\/jats:italic>, <jats:italic>IL1B<\/jats:italic>, <jats:italic>IL6<\/jats:italic>, <jats:italic>IL8<\/jats:italic>, <jats:italic>MUC5AC<\/jats:italic>, and <jats:italic>MUC5B<\/jats:italic>) was assessed by qPCR.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>Similar to parental cultures, baseline cytokine expressions were higher in pediatric OM lines than in HMEEC\u20101 and PCI, and HMEEC\u20101 cells were less responsive to stimulation than pediatric lines.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>Immortalized MEE lines retained the inflammatory expression and responsiveness of their tissues of origin and differences between non\u2010OM versus OM and pediatric versus adult cultures, supporting their value as novel in vitro culture models for OM.<\/jats:p><\/jats:sec>","DOI":"10.1002\/lio2.1141","type":"journal-article","created":{"date-parts":[[2023,8,28]],"date-time":"2023-08-28T13:01:00Z","timestamp":1693227660000},"page":"1428-1435","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Establishment of novel immortalized middle ear cell lines as models for otitis media"],"prefix":"10.1002","volume":"8","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-6027-1409","authenticated-orcid":false,"given":"Simon","family":"Blaine\u2010Sauer","sequence":"first","affiliation":[{"name":"Department of Otolaryngology and Communication Sciences Medical College of Wisconsin  Milwaukee Wisconsin USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7045-357X","authenticated-orcid":false,"given":"Tina L.","family":"Samuels","sequence":"additional","affiliation":[{"name":"Department of Otolaryngology and Communication Sciences 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