Rat tracheal epithelial cell differentiation in vitro |
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Authors: | Liisa Kaartinen Paul Nettesheim Kenneth B Adler Scott H Randell |
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Institution: | (1) Department of Pharmacology and Toxicology, College of Veterinary Medicine, SF-00581 Helsinki, Finland;(2) Laboratory of Pulmonary Pathobiology, National Institute of Environmental Health Sciences, 27709 Research Triangle Park, North Carolina;(3) Department of Anatomy, Physiological Sciences and Radiology, North Carolina State University, College of Veterinary Medicine, 27606 Raleigh, North Carolina |
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Abstract: | Summary In vitro culture conditions enabling rat tracheal epithelial (RTE) cells to differentiate to mucociliary, mucous, or squamous
phenotypes are described. Medium composition for rapid cell growth to confluence in membrane insert cultures was determined,
and the effects of major modifiers of differentiation were tested. Retinoic acid (RA), collagen gel substratum, and an air-liquid
interface at the level of the cell layer were required for expression of a mucociliary phenotype which most closely approximated
the morphology of the tracheal epithelium in vivo. Large quantities of high molecular weight, hyaluronidase-resistant glycoconjugates,
most likely mucin glycoproteins, were produced in the presence of RA when the cells were grown with or without a collagen
gel and in submerged as well as in interface cultures. However, extensive ciliagenesis was dependent on the simultaneous presence
of RA, collagen gel, and an air-liquid interface. When RA was omitted from the media, the cells became stratified squamous
and developed a cornified apical layer in air-liquid interface cultures. This phenotype was accompanied by loss of transglutaminase
(TGase) type II and keratin 18 and expression of the squamous markers TGase type I and keratin 13. The ability to modulate
RTE cell phenotypes in culture will facilitate future studies investigating molecular regulation of tracheal cell proliferation,
differentiation, and function. |
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Keywords: | collagen differentiation keratin mucin retinoids transglutaminase |
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