title: The characterization of the human nasal epithelial cell line RPMI 2650 under different culture conditions and their optimization for an appropriate in vitro nasal model.
abstract: OBJECTIVE
  The further characterization of the cell line RPMI 2650 and the evaluation of different culture conditions for an in vitro model for nasal mucosa.
  
  
  METHODS
  Cells were cultured in media MEM or A-MEM at air-liquid (A-L) or liquid-liquid (L-L) interfaces for 1 or 3 weeks. Different cryopreservation methods and cell culture techniques were evaluated with immunolabelling of junctional proteins, ultrastructural analysis using electron microscopy, transepithelial electrical resistance (TEER) measurements, permeation studies with dextran and jacalin, and gene expression profiling of 84 drug transporters.
  
  
  RESULTS
  Cell proliferation and differentiation depended on the used medium. The established epithelia expressed occludin, claudin-1, and E-cadherin under all conditions. Cells grown at the A-L interface formed more layers and exhibited a higher TEER and lower dextran and jacalin permeability than at the L-L interface, where cells morphologically exhibited a more differentiated phenotype. The expression of ABC and SLC transporters depended on culture duration and interface.
  
  
  CONCLUSIONS
  The RPMI 2650 cells form a polarized epithelium resembling nasal mucosa. However, different culture conditions have a significant effect on cell ultrastructure, barrier integrity, and gene expression, and should be considered when using this cell line as an in vitro model for drug permeability studies and screening of nasal drug candidates.
authors.affiliation: Institute of Cell Biology, Faculty of Medicine, University of Ljubljana, Ljubljana, Slovenia.
authors.forename: Mateja Erdani
authors.initials: ME
authors.lastname: Kreft
authors.forename: Urška Dragin
authors.initials: UD
authors.lastname: Jerman
authors.forename: Eva
authors.initials: E
authors.lastname: Lasič
authors.forename: Tea
authors.initials: T
authors.lastname: Lanišnik Rižner
authors.forename: Neli
authors.initials: N
authors.lastname: Hevir-Kene
authors.forename: Luka
authors.initials: L
authors.lastname: Peternel
authors.forename: Katja
authors.initials: K
authors.lastname: Kristan
mesh_terms: D018929:Cell Culture Techniques / Q000379:methods*; D002460:Cell Line; D049109:Cell Proliferation / Q000502:physiology; D002470:Cell Survival / Q000502:physiology; D006801:Humans; D008954:Models, Biological; D009297:Nasal Mucosa / Q000166:cytology* / Q000378:metabolism* / Q000648:ultrastructure