Isolation of a putative progenitor subpopulation of alveolar epithelial type 2 cells |
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Authors: | Reddy Raghava Buckley Sue Doerken Melissa Barsky Lora Weinberg Kenneth Anderson Kathryn D Warburton David Driscoll Barbara |
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Institution: | Childrens Hospital Los Angeles Research Institute, Smith Research Towe, MS 35, 4650 Sunset Blvd., Los Angeles, CA 90027, USA. |
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Abstract: | 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, -, - and -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. |
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