title: Growth regulation of serum-free cultures of epithelial cells from normal human buccal mucosa.
abstract: Human buccal epithelial cells have been reared from explants maintained in supplemented MCDB 153 medium. Primary epithelial outgrowths show typical structural features and uniformly express keratins; subunit analyses demonstrate expression of keratins 5, 6, 14, 16/17, and 19. The cells exhibit up to 6% colony forming efficiency and divide at about 0.8 population doublings per day on fibronectin/collagen-coated dishes at clonal density. Studies of markers of proliferation and differentiation in buccal epithelial cells indicate that epidermal growth factor, cholera toxin, retinoic acid, and pituitary extract each exhibit a distinctive ability to enhance growth and variably affect cell migration and cell surface area. Transforming growth factor beta-1 inhibits growth and increases surface area without affecting migration, involucrin expression, and cross-linked envelope formation. Moreover, exposure of cells to fetal bovine serum, the tumor promoting agent 12-O-tetradecanoylphorbol-13-acetate or an elevated Ca2+ concentration (from 0.1 to 1 mM) inhibits growth and induces squamous differentiation as indicated by inhibition of migration, increases in surface area, involucrin expression, or formation of cross-linked envelopes. The results show that epithelial cells can be reproducibly derived from explant cultures of human buccal mucosa specimens and the cells transferred under serum-free conditions. Buccal epithelial cells in culture undergo a pattern of growth and differentiation that mimics parakeratinization in vivo and variably respond to several agents shown to modulate growth of cells that originate from other types of epithelia.
authors.affiliation: Department of Toxicology, Karolinska Institutet, Stockholm, Sweden.
authors.forename: K
authors.initials: K
authors.lastname: Sundqvist
authors.forename: Y
authors.initials: Y
authors.lastname: Liu
authors.forename: K
authors.initials: K
authors.lastname: Arvidson
authors.forename: K
authors.initials: K
authors.lastname: Ormstad
authors.forename: L
authors.initials: L
authors.lastname: Nilsson
authors.forename: R
authors.initials: R
authors.lastname: Toftgård
authors.forename: R C
authors.initials: RC
authors.lastname: Grafström
mesh_terms: D000818:Animals; D001769:Blood; D002118:Calcium / Q000494:pharmacology; D002417:Cattle; D002455:Cell Division; D002772:Cholera Toxin / Q000494:pharmacology; D003470:Culture Media; D004815:Epidermal Growth Factor / Q000494:pharmacology; D004847:Epithelial Cells; D004848:Epithelium / Q000378:metabolism; D006801:Humans; D066298:In Vitro Techniques; D007633:Keratins / Q000378:metabolism; D008855:Microscopy, Electron, Scanning; D009061:Mouth Mucosa / Q000166:cytology* / Q000378:metabolism; D010907:Pituitary Hormones / Q000494:pharmacology; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D016212:Transforming Growth Factor beta / Q000494:pharmacology; D014212:Tretinoin / Q000494:pharmacology
chemical_list: D003470:Culture Media; D010907:Pituitary Hormones; D016212:Transforming Growth Factor beta; D014212:Tretinoin; D004815:Epidermal Growth Factor; D007633:Keratins; D002772:Cholera Toxin; D013755:Tetradecanoylphorbol Acetate; D002118:Calcium