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Am J Physiol Lung Cell Mol Physiol 286: L658-L667, 2004. First published August 15, 2003; doi:10.1152/ajplung.00159.2003
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Stem Cells in Lung Biology

Isolation of a putative progenitor subpopulation of alveolar epithelial type 2 cells

Raghava Reddy, Sue Buckley, Melissa Doerken, Lora Barsky, Kenneth Weinberg, Kathryn D. Anderson, David Warburton, and Barbara Driscoll

Department of Surgery and Developmental Biology Program, Childrens Hospital Los Angeles Research Institute, University of Southern California School of Medicine, Los Angeles, California 90027

Submitted 20 May 2003 ; accepted in final form 8 August 2003

Alveolar epithelial type 2 cells (AEC2) isolated from hyperoxia-treated animals exhibit increases in both proliferation and DNA damage in response to culture. AEC2 express the zonula adherens proteins E-cadherin, {alpha}-, {beta}- and {gamma}-catenin, desmoglein, and pp120, as demonstrated by Western blotting. Immunohistochemical analysis of cultured AEC2 showed expression of E-cadherin on cytoplasmic membranes varying from strongly to weakly staining. When cultured AEC2 placed in suspension were labeled with fluorescent-tagged antibodies to E-cadherin, cells could be sorted into at least two subpopulations, either dim or brightly staining for this marker. With the use of antibody to E-cadherin bound to magnetic beads, cells were physically separated into E-cadherin-positive and -negative subpopulations, which were then analyzed for differences in proliferation and DNA damage. The E-cadherin-positive subpopulation contained the majority of damaged cells, was quiescent, and expressed low levels of telomerase activity, whereas the E-cadherin-negative subpopulation was undamaged, proliferative, and expressed high levels of telomerase activity.

alveolar epithelial cells; hyperoxic injury; E-cadherin; proliferation


Address for reprint requests and other correspondence: B. Driscoll, Childrens Hospital Los Angeles Research Institute, Smith Research Tower, MS 35, 4650 Sunset Blvd., Los Angeles, CA 90027 (E-mail: bdriscoll{at}chla.usc.edu).




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