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Published ahead of print on December 2, 2004, doi:10.1165/rcmb.2004-0079RC

Am. J. Respir. Cell Mol. Biol., Volume 32, Number 2, February 2005, 87-92

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Submitted on March 1, 2004
Revised on December 1, 2004

EMBRYONIC STEM CELLS GENERATE AIRWAY EPITHELIAL TISSUE

Christelle Coraux1, Beatrice Nawrocki-Raby1, Jocelyne Hinnrasky1, Claire Kileztky1, Dominique Gaillard1, Christian Dani2, and Edith Puchelle1*

1 INSERM UMR S 514 CHU Maison Blanche, Reims, France, 2 CNRS UMR 6543, Centre de Biochimie, Nice, France

* To whom correspondence should be addressed. E-mail: edith.puchelle{at}univ-reims.fr.

Embryonic stem (ES) cells are self-renewable and pluripotent cells derived from the inner cell mass of a blastocyst-stage embryo. ES cell pluripotency is being investigated increasingly to obtain specific cell lineages for therapeutic treatments and tissue engineering. Type II alveolar epithelial cells have been derived from murine ES cells, but the capacity of the latter to generate differentiated airway epithelial tissue has never been reported. Herein, we show by RT-PCR and immunocytochemistry that murine ES cells are able to differentiate into nonciliated secretory Clara cells, and that type I collagen induces this commitment. Moreover, when cultured at the air-liquid interface, ES cells give rise to a fully differentiated airway epithelium. By quantitative histological examination, immunohistochemistry and scanning electron microscopy, we show that the bioengineered epithelium is composed of basal, ciliated, intermediate and Clara cells, similar to those of native tracheobronchial airway epithelium. Transmission electron microscopy and western blotting reveal that the generated epithelium also exhibits the ultrastructural features and secretory functions characteristic of airway epithelial tissue. These results open new perspectives for cell therapy of injured epithelium in airway diseases, such as bronchopulmonary dysplasia, cystic fibrosis or bronchiolitis obliterans.




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