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Related Experiment Videos

Side population cells and Bcrp1 expression in lung.

Ross Summer1, Darrell N Kotton, Xi Sun

  • 1The Pulmonary Center, Boston University School of Medicine, 80 E. Concord St., Boston, MA 02118, USA. rsummer@lung.bumc.bu.edu

American Journal of Physiology. Lung Cellular and Molecular Physiology
|March 11, 2003
PubMed
Summary

Side population (SP) cells, enriched for stem cell activity, were identified in mouse lungs using Hoechst dye efflux. These rare cells express BCRP1, found in smooth muscle and airway cells, influencing their stem cell phenotype.

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Area of Science:

  • Cell Biology
  • Stem Cell Research
  • Respiratory Medicine

Background:

  • Side population (SP) cells are rare, stem cell-enriched cells identified by their ability to efflux Hoechst dye.
  • This efflux is mediated by the ATP-binding cassette transporter breast cancer resistance protein (Bcrp) 1.
  • Understanding SP cell localization and function in the lung is crucial for regenerative medicine.

Purpose of the Study:

  • To identify and characterize side population (SP) cells in the mouse lung.
  • To investigate the role of breast cancer resistance protein 1 (Bcrp1) in lung SP cell phenotype.
  • To determine the in vivo localization of Bcrp1-expressing cells in the lung.

Main Methods:

  • Flow cytometry of enzymedigested mouse lung stained with Hoechst dye.

Related Experiment Videos

  • RT-PCR for gene expression analysis (HNF-3beta, TTF-1).
  • Surface marker analysis (Sca-1, Bcrp1, lineage markers, CD45).
  • Nonisotopic in situ hybridization and immunostaining for Bcrp1 localization.
  • Primary bronchial smooth muscle cell (BSMC) and distal airway cell culture.

    • Main Results:

    • Lung SP cells constitute 0.03-0.07% of total lung cells and are evenly distributed.
    • Lung SP cells express stem cell antigen 1 (Sca-1), Bcrp1, and are lineage marker negative.
    • Bcrp1 expression was observed in bronchial smooth muscle cells, vascular smooth muscle cells, and distal air space cells.
    • Primary BSMC and distal airway cells expressed Bcrp1 but lacked Hoechst efflux capacity.
    • Bcrp1 localization within an intracellular compartment in BSMC suggests regulation of SP phenotype.

    Conclusions:

    • Mouse lung contains a distinct side population (SP) of stem cell-enriched cells.
    • Breast cancer resistance protein 1 (Bcrp1) is critical for the SP phenotype in lung cells.
    • Bcrp1 expression in specific lung cell types, particularly smooth muscle cells, is linked to SP cell characteristics.
    • Intracellular localization of Bcrp1 influences the SP phenotype, highlighting the importance of transporter localization.