Characterization of side population cells from human airway epithelium

Tillie Louise Hackett, Furquan Shaheen, Andrew Johnson, Samuel Wadsworth, Dmitri V. Pechkovsky, David Jacoby, Anthony Kicic, Stephen M. Stick, Darryl A. Knight

Research output: Contribution to journalArticle

96 Citations (Scopus)

Abstract

The airway epithelium is the first line of contact with the inhaled external environment and is continuously exposed to and injured by pollutants, allergens, and viruses. However, little is known about epithelial repair and in particular the identity and role of tissue resident stem/progenitor cells that may contribute to epithelial regeneration. The aims of the present study were to identify, isolate, and characterize side population (SP) cells in human tracheobronchial epithelium. Epithelial cells were obtained from seven nontransplantable healthy lungs and four asthmatic lungs by pronase digestion. SP cells were identified by verapamil-sensitive efflux of the DNA-binding dye Hoechst 33342. Using flow cytometry, CD45- SP, CD45+ SP, and non-SP cells were isolated and sorted. CD45- SP cells made up 0.12% ± 0.01% of the total epithelial cell population in normal airway but 4.1% ± 0.06% of the epithelium in asthmatic airways. All CD45 - SP cells showed positive staining for epithelial-specific markers cytokeratin-5, E-cadherin, ZO-1, and p63. CD45- SP cells exhibited stable telomere length and increased colony-forming and proliferative potential, undergoing population expansion for at least 16 consecutive passages. In contrast with non-SP cells, fewer than 100 CD45- SP cells were able to generate a multilayered and differentiated epithelium in air-liquid interface culture. SP cells are present in human tracheobronchial epithelium, exhibit both short- and long-term proliferative potential, and are capable of generation of differentiated epithelium in vitro. The number of SP cells is significantly greater in asthmatic airways, providing evidence of dysregulated resident SP cells in the asthmatic epithelium.

Original languageEnglish (US)
Pages (from-to)2576-2585
Number of pages10
JournalStem Cells
Volume26
Issue number10
DOIs
StatePublished - Oct 2008

Fingerprint

Side-Population Cells
Epithelium
Population
Cadherins
Stem Cells
Epithelial Cells
Keratin-5
Lung
Pronase
Telomere
Verapamil
Allergens
Regeneration
Digestion
Flow Cytometry
Coloring Agents
Air
Staining and Labeling

Keywords

  • Asthma
  • Epithelium
  • Human
  • Tissue-specific stem cell

ASJC Scopus subject areas

  • Cell Biology
  • Developmental Biology
  • Molecular Medicine

Cite this

Hackett, T. L., Shaheen, F., Johnson, A., Wadsworth, S., Pechkovsky, D. V., Jacoby, D., ... Knight, D. A. (2008). Characterization of side population cells from human airway epithelium. Stem Cells, 26(10), 2576-2585. https://doi.org/10.1634/stemcells.2008-0171

Characterization of side population cells from human airway epithelium. / Hackett, Tillie Louise; Shaheen, Furquan; Johnson, Andrew; Wadsworth, Samuel; Pechkovsky, Dmitri V.; Jacoby, David; Kicic, Anthony; Stick, Stephen M.; Knight, Darryl A.

In: Stem Cells, Vol. 26, No. 10, 10.2008, p. 2576-2585.

Research output: Contribution to journalArticle

Hackett, TL, Shaheen, F, Johnson, A, Wadsworth, S, Pechkovsky, DV, Jacoby, D, Kicic, A, Stick, SM & Knight, DA 2008, 'Characterization of side population cells from human airway epithelium', Stem Cells, vol. 26, no. 10, pp. 2576-2585. https://doi.org/10.1634/stemcells.2008-0171
Hackett TL, Shaheen F, Johnson A, Wadsworth S, Pechkovsky DV, Jacoby D et al. Characterization of side population cells from human airway epithelium. Stem Cells. 2008 Oct;26(10):2576-2585. https://doi.org/10.1634/stemcells.2008-0171
Hackett, Tillie Louise ; Shaheen, Furquan ; Johnson, Andrew ; Wadsworth, Samuel ; Pechkovsky, Dmitri V. ; Jacoby, David ; Kicic, Anthony ; Stick, Stephen M. ; Knight, Darryl A. / Characterization of side population cells from human airway epithelium. In: Stem Cells. 2008 ; Vol. 26, No. 10. pp. 2576-2585.
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