Rat tracheal epithelial cell differentiation in vitro.
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.
Kaartinen
L
L
Department of Pharmacology and Toxicology, College of Veterinary Medicine, Helsinki, Finland.
Nettesheim
P
P
Adler
K B
KB
Randell
S H
SH
D000818:Animals; D000911:Antibodies, Monoclonal; D002454:Cell Differentiation / Q000502:physiology; D003094:Collagen / Q000494:pharmacology; D003470:Culture Media; D004847:Epithelial Cells; D004848:Epithelium / Q000187:drug effects / Q000378:metabolism; D007101:Immersion; D007150:Immunohistochemistry; D066298:In Vitro Techniques; D007633:Keratins / Q000378:metabolism; D008297:Male; D010641:Phenotype; D051381:Rats; D011916:Rats, Inbred F344; D014132:Trachea / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D011503:Transglutaminases / Q000378:metabolism; D014212:Tretinoin / Q000494:pharmacology
D000911:Antibodies, Monoclonal; D003470:Culture Media; D014212:Tretinoin; D007633:Keratins; D003094:Collagen; D011503:Transglutaminases