Generation of a human airway epithelium derived basal cell line with multipotent differentiation capacity

Matthew S. Walters, Kazunori Gomi, Beth Ashbridge, Malcolm A.S. Moore, Vanessa Arbelaez, Jonna Heldrich, Bi Sen Ding, Shahin Rafii, Michelle R. Staudt, Ronald Crystal

Research output: Contribution to journalArticle

34 Citations (Scopus)

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.Conclusion: 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.

Original languageEnglish
Article number135
JournalRespiratory Research
Volume14
Issue number1
DOIs
Publication statusPublished - 3 Dec 2013
Externally publishedYes

Fingerprint

Epithelium
Cell Line
Air
Aptitude
Interleukin-13
Cell Aging
Telomerase
Human Development
Retroviridae
Population
Cell Biology
Clone Cells
Enzyme-Linked Immunosorbent Assay
Staining and Labeling
Phenotype
Polymerase Chain Reaction

Keywords

  • Airway
  • Basal cell
  • Differentiation
  • hTERT
  • Immortalized
  • Progenitor

ASJC Scopus subject areas

  • Pulmonary and Respiratory Medicine

Cite this

Generation of a human airway epithelium derived basal cell line with multipotent differentiation capacity. / Walters, Matthew S.; Gomi, Kazunori; Ashbridge, Beth; Moore, Malcolm A.S.; Arbelaez, Vanessa; Heldrich, Jonna; Ding, Bi Sen; Rafii, Shahin; Staudt, Michelle R.; Crystal, Ronald.

In: Respiratory Research, Vol. 14, No. 1, 135, 03.12.2013.

Research output: Contribution to journalArticle

Walters, MS, Gomi, K, Ashbridge, B, Moore, MAS, Arbelaez, V, Heldrich, J, Ding, BS, Rafii, S, Staudt, MR & Crystal, R 2013, 'Generation of a human airway epithelium derived basal cell line with multipotent differentiation capacity', Respiratory Research, vol. 14, no. 1, 135. https://doi.org/10.1186/1465-9921-14-135
Walters, Matthew S. ; Gomi, Kazunori ; Ashbridge, Beth ; Moore, Malcolm A.S. ; Arbelaez, Vanessa ; Heldrich, Jonna ; Ding, Bi Sen ; Rafii, Shahin ; Staudt, Michelle R. ; Crystal, Ronald. / Generation of a human airway epithelium derived basal cell line with multipotent differentiation capacity. In: Respiratory Research. 2013 ; Vol. 14, No. 1.
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AU - Gomi, Kazunori

AU - Ashbridge, Beth

AU - Moore, Malcolm A.S.

AU - Arbelaez, Vanessa

AU - Heldrich, Jonna

AU - Ding, Bi Sen

AU - Rafii, Shahin

AU - Staudt, Michelle R.

AU - Crystal, Ronald

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