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Am J Physiol Lung Cell Mol Physiol 286: L624-L630, 2004. First published August 8, 2003; doi:10.1152/ajplung.00149.2003
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Stem Cells in Lung Biology

Molecular phenotype of airway side population cells

Adam Giangreco,1,2 Hongmei Shen,3 Susan D. Reynolds,1 and Barry R. Stripp1,2

Departments of 1Environmental and Occupational Health, 2Cell Biology and Physiology, and 3Radiation Oncology, University of Pittsburgh, Pittsburgh, Pennsylvania 15260

Submitted 14 May 2003 ; accepted in final form 30 July 2003

Lung epithelial-specific stem cells have been localized to discrete microenvironments throughout the adult conducting airway. Properties of these cells include pollutant resistance, multipotent differentiation, and infrequent proliferation. Goals of the present study were to use Hoechst 33342 efflux, a property of stem cells in other tissues, to purify and further characterize airway stem cells. Hoechst 33342 effluxing lung cells were identified as a verapamil-sensitive side population by flow cytometry. Lung side population cells were further subdivided on the basis of hematopoietic (CD45 positive) or nonhematopoietic (CD45 negative) origin. Nonhematopoietic side population cells were enriched for stem cell antigen-1 reactivity and expressed molecular markers specific to both airway and mesenchymal lineages. Analysis of the molecular phenotype of airway-derived side population cells indicates that they are similar to neuroepithelial body-associated variant Clara cells. Taken together, these data suggest that the nonhematopoietic side population isolated from lung is enriched for previously identified airway stem cells.

stem cell; Clara; lung; Hoechst 33342



Address for reprint requests and other correspondence: B. R. Stripp, Univ. of Pittsburgh, FORBL Rm. 314, 3343 Forbes Ave., Pittsburgh, PA 15260 (E-mail: brs2{at}pitt.edu).




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