title: Changes in the gene expression pattern of cytokeratins in human respiratory epithelial cells during culture.
abstract: The replacement of extensive tracheal defects resulting from intensive care medicine, trauma or large resections is still challenged by the re-epithelialization of an autologous or alloplastic trachea replacement. Therefore, this study was performed to investigate the potential of culture-expanded human respiratory epithelial cells (hREC) to regenerate a functional epithelium for tracheal tissue engineering. hREC from seven male nasal turbinates were freshly isolated, expanded on a collagenous matrix and subsequently cultured in high-density multi-layers to allow epithelial differentiation. The composition of epithelial cells in native respiratory epithelial tissue and culture-expanded hREC was analyzed by histological staining with Alcian blue and by immunohistochemical staining of cytokeratin pairs CK1/10 and CK5/14 with the antibodies 34betaE12 and CD44v6. Differentiation of culture-expanded hREC was further characterized by gene expression analysis of cytokeratins CK5, CK13, CK14 and CK18 using semi-quantitative real-time RT-PCR technique. Histological and immunohistochemical staining of culture-expanded hREC demonstrated basal cells covering the collagenous matrix. These cells formed a cellular multi-layer, which was composed of a basal layer of undifferentiated basal cells and an upper layer of cells differentiating along the squamous metaplasia and ciliated cell lineage. Lineage development of culture-expanded hREC was further documented by the induction of cytokeratins CK13 and CK18. Our results suggest that culture-expanded hREC have the potential to colonize collagen-coated biomaterials and to regenerate epithelial cell types for tracheal tissue engineering.
authors.affiliation: Tissue Engineering Laboratory, University Medical Center Charité, Tucholskystr. 2, 10117 Berlin, Germany. michaela.endres@charite.de
authors.forename: Michaela
authors.initials: M
authors.lastname: Endres
authors.forename: Iris
authors.initials: I
authors.lastname: Leinhase
authors.forename: Christian
authors.initials: C
authors.lastname: Kaps
authors.forename: Marek
authors.initials: M
authors.lastname: Wentges
authors.forename: Manja
authors.initials: M
authors.lastname: Unger
authors.forename: Heidi
authors.initials: H
authors.lastname: Olze
authors.forename: Jochen
authors.initials: J
authors.lastname: Ringe
authors.forename: Michael
authors.initials: M
authors.lastname: Sittinger
authors.forename: Nicole
authors.initials: N
authors.lastname: Rotter
mesh_terms: D000328:Adult; D015415:Biomarkers / Q000032:analysis; D004847:Epithelial Cells / Q000166:cytology* / Q000378:metabolism; D015870:Gene Expression / Q000235:genetics*; D006801:Humans; D007150:Immunohistochemistry; D007633:Keratins / Q000096:biosynthesis* / Q000235:genetics; D008297:Male; D008875:Middle Aged; D016133:Polymerase Chain Reaction; D020545:Respiratory Mucosa / Q000166:cytology*; D014420:Turbinates / Q000378:metabolism
chemical_list: D015415:Biomarkers; D007633:Keratins