title: Alternative Method for Primary Nasal Epithelial Cell Culture Using Intranasal Brushing and Feasibility for the Study of Epithelial Functions in Allergic Rhinitis.
abstract: OBJECTIVE
  Although differentiated normal human nasal epithelial (NHNE) cells can be used to study the role of human nasal epithelium, there is a need for effective culture models of nasal epithelium in sinonasal disease status, including allergic rhinitis (AR). We aimed to examine the feasibility of intranasal brushing for culture of nasal epithelial cells in AR patients and to verify the hypothesis that allergic nasal epithelial (ARNE) cells differ in histologic and physiologic characteristics.
  
  
  METHODS
  We established a system for isolating (via intranasal brushing) and culturing (with air-liquid interface, ALI) nasal epithelial cells from healthy volunteers (n=8) and AR patients (n=8). We used this system to compare the histologic findings and physiologic characteristics of NHNE and ARNE.
  
  
  RESULTS
  The histology results showed that fully differentiated ALI culture was obtained at least 14 days after confluence and that both ciliated and secretory cells were well differentiated in ALI culture using nasal brushing. The histology results of ARNE culture were significantly different from NHNE. The number of ciliated cells was lower, and secretory cells were more dominant in ARNE cell culture compared to NHNE cells. We also observed, by electron microscopy, loose tight junctions and short cilia in cultured ARNE cells. In addition, the mRNA level of TSLP which was one of the epithelial-derived allergic cytokines was significantly higher, and the expressions of genes involved in ciliogenesis were lower in cultured ARNE cells without allergen stimulation.
  
  
  CONCLUSIONS
  Our findings suggest that ALI culture of ARNE cells using intranasal brushing may be an alternative method for epithelial cell culture in AR patients and that cultured ARNE cells will be useful for in vitro studies of the mechanisms at play during AR because they maintain unique allergic characteristics.
authors.affiliation: Department of Otolaryngology-Head and Neck Surgery, Soonchunhyang University College of Medicine, Cheonan Hospital, Cheonan, Korea.
authors.forename: Do Yang
authors.initials: DY
authors.lastname: Park
authors.affiliation: BK21 PLUS Project for Medical Science, Yonsei University College of Medicine, Seoul, Korea.
authors.forename: Sujin
authors.initials: S
authors.lastname: Kim
authors.affiliation: Department of Otorhinolaryngology, Yonsei University College of Medicine, Seoul, Korea.|Airway Mucus Institute, Yonsei University College of Medicine, Seoul, Korea.|Research Center for Natural Human Defense System, Yonsei University College of Medicine, Seoul, Korea.
authors.forename: Chang Hoon
authors.initials: CH
authors.lastname: Kim
authors.affiliation: Department of Otorhinolaryngology, Yonsei University College of Medicine, Seoul, Korea.|Airway Mucus Institute, Yonsei University College of Medicine, Seoul, Korea.|Research Center for Natural Human Defense System, Yonsei University College of Medicine, Seoul, Korea.
authors.forename: Joo Heon
authors.initials: JH
authors.lastname: Yoon
authors.affiliation: Airway Mucus Institute, Yonsei University College of Medicine, Seoul, Korea.|Department of Otorhinolaryngology, Seoul National University College of Medicine, Seoul, Korea. hyunjerry@snu.ac.kr.
authors.forename: Hyun Jik
authors.initials: HJ
authors.lastname: Kim
keywords: Intranasal brushing; allergic rhinitis; human nasal epithelial cells; primary cell culture