title: An in vitro model of murine middle ear epithelium.
abstract: Otitis media (OM), or middle ear inflammation, is the most common paediatric disease and leads to significant morbidity. Although understanding of underlying disease mechanisms is hampered by complex pathophysiology it is clear that epithelial abnormalities underpin the disease. There is currently a lack of a well-characterised in vitro model of the middle ear (ME) epithelium that replicates the complex cellular composition of the middle ear. Here, we report the development of a novel in vitro model of mouse middle ear epithelial cells (mMECs) at an air-liquid interface (ALI) that recapitulates the characteristics of the native murine ME epithelium. We demonstrate that mMECs undergo differentiation into the varied cell populations seen within the native middle ear. Proteomic analysis confirmed that the cultures secrete a multitude of innate defence proteins from their apical surface. We showed that the mMECs supported the growth of the otopathogen, nontypeable Haemophilus influenzae (NTHi), suggesting that the model can be successfully utilised to study host-pathogen interactions in the middle ear. Overall, our mMEC culture system can help to better understand the cell biology of the middle ear and improve our understanding of the pathophysiology of OM. The model also has the potential to serve as a platform for validation of treatments designed to reverse aspects of epithelial remodelling that underpin OM development.
authors.affiliation: Academic Unit of Respiratory Medicine, Department of Infection, Immunity and Cardiovascular Disease, University of Sheffield, Sheffield S10 2JF, UK.
authors.forename: Apoorva
authors.initials: A
authors.lastname: Mulay
authors.affiliation: Academic Unit of Respiratory Medicine, Department of Infection, Immunity and Cardiovascular Disease, University of Sheffield, Sheffield S10 2JF, UK.
authors.forename: Khondoker M
authors.initials: KM
authors.lastname: Akram
authors.affiliation: MRC Mammalian Genetics Unit, Harwell OX11 0RD, UK.
authors.forename: Debbie
authors.initials: D
authors.lastname: Williams
authors.affiliation: Academic Unit of Respiratory Medicine, Department of Infection, Immunity and Cardiovascular Disease, University of Sheffield, Sheffield S10 2JF, UK.|Oral and Maxillofacial Pathology, Department of Clinical Dentistry, University of Sheffield, Sheffield S10 2TA, UK.
authors.forename: Hannah
authors.initials: H
authors.lastname: Armes
authors.affiliation: Academic Unit of Respiratory Medicine, Department of Infection, Immunity and Cardiovascular Disease, University of Sheffield, Sheffield S10 2JF, UK.
authors.forename: Catherine
authors.initials: C
authors.lastname: Russell
authors.affiliation: MRC Mammalian Genetics Unit, Harwell OX11 0RD, UK.
authors.forename: Derek
authors.initials: D
authors.lastname: Hood
authors.affiliation: Department of Infection Biology, Institute of Infection and Global Health, University of Liverpool, Liverpool L3 5RF, UK.
authors.forename: Stuart
authors.initials: S
authors.lastname: Armstrong
authors.affiliation: Department of Infection Biology, Institute of Infection and Global Health, University of Liverpool, Liverpool L3 5RF, UK.
authors.forename: James P
authors.initials: JP
authors.lastname: Stewart
authors.affiliation: MRC Mammalian Genetics Unit, Harwell OX11 0RD, UK.
authors.forename: Steve D M
authors.initials: SD
authors.lastname: Brown
authors.affiliation: Oral and Maxillofacial Pathology, Department of Clinical Dentistry, University of Sheffield, Sheffield S10 2TA, UK.
authors.forename: Lynne
authors.initials: L
authors.lastname: Bingle
authors.affiliation: Academic Unit of Respiratory Medicine, Department of Infection, Immunity and Cardiovascular Disease, University of Sheffield, Sheffield S10 2JF, UK c.d.bingle@sheffield.ac.uk.
authors.forename: Colin D
authors.identifier: 0000-0002-5405-6988
authors.initials: CD
authors.lastname: Bingle
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D018929:Cell Culture Techniques; D048430:Cell Shape; D002478:Cells, Cultured; D002923:Cilia / Q000378:metabolism; D004432:Ear, Middle / Q000033:anatomy & histology*; D004847:Epithelial Cells / Q000378:metabolism / Q000648:ultrastructure; D004848:Epithelium / Q000033:anatomy & histology* / Q000378:metabolism / Q000648:ultrastructure; D006192:Haemophilus Infections / Q000382:microbiology / Q000473:pathology; D006193:Haemophilus influenzae / Q000502:physiology; D013058:Mass Spectrometry; D008810:Mice, Inbred C57BL; D023421:Models, Animal; D020543:Proteome / Q000378:metabolism
chemical_list: D015415:Biomarkers; D020543:Proteome
keywords: Air–liquid interface; Bpifa1; Middle ear epithelium; NTHi; Otitis media