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Am J Physiol Lung Cell Mol Physiol (March 7, 2003). doi:10.1152/ajplung.00009.2003
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Submitted on January 10, 2003
Accepted on March 5, 2003

SP (Side Population) Cells and Bcrp1 Expression in Lung

Ross Summer1*, Darrell N. Kotton1, Xi Sun1, Bei Ma1, Kathleen Fitzsimmons1, and Alan Fine1

1 Department of Pulmonary Center, Boston University School of Medicine, Boston, Massachusetts, USA

* To whom correspondence should be addressed. E-mail: rsummer{at}lung.bumc.bu.edu.

Side population (SP) cells are a rare subset of cells found in various tissues that are highly enriched for stem cell activity. SP cells can be isolated by dual-wavelength flow cytometry due to their capacity to efflux Hoechst dye, a process mediated by the ABC transporter Bcrp1. By performing flow cytometry of enzyme digested mouse lung stained with Hoechst dye, we found that SP cells comprise 0.03-0.07% of total lung cells, and are evenly distributed in proximal and distal lung regions. By RT-PCR, we found that lung SP cells express HNF-3{beta}, but not TTF-1. Surface marker analysis revealed lung SP cells to be Sca1+, Bcrp1+, lin-, and heterogeneous at the CD45 locus. As expected, we did not detect lung SP cells in Bcrp1 deficient animals. We, therefore, employed non-isotopic in situ hybridization and immuno-staining for Bcrp1 as a strategy to localize these cells in vivo. Expression was observed in distinct lung cell types: bronchial and vascular smooth muscle cells, and round cells within the distal airspace. We confirmed the expression of Bcrp1 in primary bronchial smooth muscle cell cultures (BSMC), and in lavaged distal airway cells, but neither possessed the capacity to efflux Hoechst dye. In BSMC, Bcrp1 was localized to an intracellular compartment, suggesting that the molecular site of Bcrp1 expression regulates SP phenotype.




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