title: Generation of a human airway epithelium derived basal cell line with multipotent differentiation capacity.
abstract: BACKGROUND
  As the multipotent progenitor population of the airway epithelium, human airway basal cells (BC) replenish the specialized differentiated cell populations of the mucociliated airway epithelium during physiological turnover and repair. Cultured primary BC divide a limited number of times before entering a state of replicative senescence, preventing the establishment of long-term replicating cultures of airway BC that maintain their original phenotype.
  
  
  METHODS
  To generate an immortalized human airway BC cell line, primary human airway BC obtained by brushing the airway epithelium of healthy nonsmokers were infected with a retrovirus expressing human telomerase (hTERT). The resulting immortalized cell line was then characterized under non-differentiating and differentiating air-liquid interface (ALI) culture conditions using ELISA, TaqMan quantitative PCR, Western analysis, and immunofluorescent and immunohistochemical staining analysis for cell type specific markers. In addition, the ability of the cell line to respond to environmental stimuli under differentiating ALI culture was assessed.
  
  
  RESULTS
  We successfully generated an immortalized human airway BC cell line termed BCi-NS1 via expression of hTERT. A single cell derived clone from the parental BCi-NS1 cells, BCi-NS1.1, retains characteristics of the original primary cells for over 40 passages and demonstrates a multipotent differentiation capacity into secretory (MUC5AC, MUC5B), goblet (TFF3), Clara (CC10) and ciliated (DNAI1, FOXJ1) cells on ALI culture. The cells can respond to external stimuli such as IL-13, resulting in alteration of the normal differentiation process.
  
  
  CONCLUSIONS
  Development of immortalized human airway BC that retain multipotent differentiation capacity over long-term culture should be useful in understanding the biology of BC, the response of BC to environmental stress, and as a target for assessment of pharmacologic agents.
authors.forename: Matthew S
authors.initials: MS
authors.lastname: Walters
authors.forename: Kazunori
authors.initials: K
authors.lastname: Gomi
authors.forename: Beth
authors.initials: B
authors.lastname: Ashbridge
authors.forename: Malcolm A S
authors.initials: MA
authors.lastname: Moore
authors.forename: Vanessa
authors.initials: V
authors.lastname: Arbelaez
authors.forename: Jonna
authors.initials: J
authors.lastname: Heldrich
authors.forename: Bi-Sen
authors.initials: BS
authors.lastname: Ding
authors.forename: Shahin
authors.initials: S
authors.lastname: Rafii
authors.forename: Michelle R
authors.initials: MR
authors.lastname: Staudt
authors.affiliation: Department of Genetic Medicine, Weill Cornell Medical College, New York NY, USA. geneticmedicine@med.cornell.edu.
authors.forename: Ronald G
authors.initials: RG
authors.lastname: Crystal
mesh_terms: D000328:Adult; D001980:Bronchi / Q000166:cytology* / Q000378:metabolism; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation; D002460:Cell Line; D004847:Epithelial Cells / Q000166:cytology* / Q000378:metabolism; D006801:Humans; D008297:Male; D039902:Multipotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D010641:Phenotype; D012190:Retroviridae / Q000235:genetics; D019098:Telomerase / Q000235:genetics / Q000378:metabolism; D014162:Transfection
chemical_list: C509186:TERT protein, human; D019098:Telomerase