Establishment of novel immortalized middle ear cell lines as models for otitis media. Laryngoscope Investig Otolaryngol 2023 Oct;8(5):1428-1435
Date
10/30/2023Pubmed ID
37899851Pubmed Central ID
PMC10601576DOI
10.1002/lio2.1141Scopus ID
2-s2.0-85168915314 (requires institutional sign-in at Scopus site) 2 CitationsAbstract
OBJECTIVE: 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-1, adult-derived). We previously developed MEE cultures from pediatric patients with non-inflamed 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-relevant pathogen lysate and cytokines. Herein, we sought to immortalize these cultures and assess retention of their phenotypes.
METHODS: MEE cultures were immortalized via lentivirus encoding temperature-sensitive SV40 T antigen. Immortalized MEE lines and HMEEC-1 grown in monolayer were stimulated with non-typeable Haemophilus influenzae (NTHi) lysate. Gene expression (TNFA, IL1B, IL6, IL8, MUC5AC, and MUC5B) was assessed by qPCR.
RESULTS: Similar to parental cultures, baseline cytokine expressions were higher in pediatric OM lines than in HMEEC-1 and PCI, and HMEEC-1 cells were less responsive to stimulation than pediatric lines.
CONCLUSION: Immortalized MEE lines retained the inflammatory expression and responsiveness of their tissues of origin and differences between non-OM versus OM and pediatric versus adult cultures, supporting their value as novel in vitro culture models for OM.
Author List
Blaine-Sauer S, Samuels TL, Khampang P, Yan K, McCormick ME, Chun RH, Harvey SA, Friedland DR, Johnston N, Kerschner JEAuthors
Robert H. Chun MD Professor in the Otolaryngology department at Medical College of WisconsinDavid R. Friedland MD Associate Director, Director, Chief, Professor in the Otolaryngology department at Medical College of Wisconsin
Steven A. Harvey MD Associate Professor in the Otolaryngology department at Medical College of Wisconsin
Nikki Johnston PhD Professor in the Otolaryngology department at Medical College of Wisconsin
Michael E. McCormick MD Associate Professor in the Otolaryngology department at Medical College of Wisconsin
