ArticleLaryngoscope investigative otolaryngology2019
Human primary middle ear epithelial cell culture: A novel in vitro model to study otitis media.
Article in Laryngoscope investigative otolaryngology, 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 6 papers.
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Who cites it
6 citing papers in PubMed, 10 citations in OpenAlex.
- Advances and prospects of 3D air-liquid interface cultured middle ear epithelial cells and inner ear organoids in otologic diseases.Frontiers in bioengineering and biotechnology · 2026Review
- Establishment of novel immortalized middle ear cell lines as models for otitis media.Laryngoscope investigative otolaryngology · 2023Article
- Nebulized Menthol Impairs Mucociliary Clearance via TRPM8 and MUC5AC/MUC5B in Primary Airway Epithelial Cells.International journal of molecular sciences · 2023Article
- Development and Characterization of an In Vitro Round Window Membrane Model for Drug Permeability Evaluations.Pharmaceuticals (Basel, Switzerland) · 2022Article
- Development of a physiological model of human middle ear epithelium.Laryngoscope investigative otolaryngology · 2021Article
- The transcriptional landscape of the cultured murine middle ear epithelium in vitro.Biology open · 2021Article
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Authors and funding
8 authors at 1 institution in 1 country.
Funding
Abstract
objectivesOtitis media (OM) is a ubiquitous pediatric disease leading to a significant health care burden. There is no medication beneficial to resolving COM fluid, highlighting the need for research in the field. Crucially, current human middle ear epithelial cell models are transformed cells not recapitulating physiological functions. Herein, we describe a new method to proliferate and differentiate pediatric primary middle ear epithelial cells (pMEEC) from patients as a physiological model for the study of OM.
methodsWe adapted a cell reprogramming protocol using irradiated fibroblast feeder medium in addition to Rho kinase inhibitor to proliferate pMEEC collected during cochlear implant surgery. Cells were plated on transwell membranes, proliferated with conditionally reprogrammed culture medium, and transferred to air-liquid interface (ALI). Cultures were maintained for 4 weeks at ALI, photos were taken and cell lysates and secretions were collected over time for characterization analysis using quantitative polymerase chain reaction, Western bolt, and proteomics. Keratins, MUC5B and MUC5AC mucins, and beta tubulin (TUBB) were analyzed at the mRNA and protein level.
resultsCultures took a mean of 2 weeks to proliferate before transwell plating and forming a tight epithelium at ALI from 2 to 4 weeks. Although mRNA expression of MUC5B, MUC5AC, TUBB, and keratin 5 (KRT5) were variable depending on the differentiation stage and the patient, both TUBB and KRT5 proteins were detected until week 2.
conclusionWe demonstrate a novel method to proliferate and differentiate pMEECs that express epithelial markers and that are able to secrete mucins for the study of OM. LEVEL OF EVIDENCE: NA.
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