Generation of a human airway epithelium derived basal cell line with multipotent differentiation capacity. Academic Article uri icon

Overview

MeSH

  • Adult
  • Cell Line
  • Humans
  • Male
  • Phenotype
  • Retroviridae
  • Telomerase
  • Transfection

MeSH Major

  • Bronchi
  • Cell Culture Techniques
  • Cell Differentiation
  • Epithelial Cells
  • Multipotent Stem Cells

abstract

  • 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. 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. 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. 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.

publication date

  • December 3, 2013

has subject area

  • Adult
  • Bronchi
  • Cell Culture Techniques
  • Cell Differentiation
  • Cell Line
  • Epithelial Cells
  • Humans
  • Male
  • Multipotent Stem Cells
  • Phenotype
  • Retroviridae
  • Telomerase
  • Transfection

Research

keywords

  • Journal Article

Identity

Language

  • eng

PubMed Central ID

  • PMC3907041

Digital Object Identifier (DOI)

  • 10.1186/1465-9921-14-135

PubMed ID

  • 24298994

Additional Document Info

start page

  • 135

volume

  • 14