Growth regulation of serum-free cultures of epithelial cells from normal human buccal mucosa

• DOI: 10.1007/bf02631287
• Publication Date: 2007-05-19T17:46:25Z
• Authors: Kristina Sundqvist; Yun Liu; Kristina Arvidson; Kari Ormstad; Lennart Nilsson; Rune Toftgård; Roland C. Grafström
• Keywords: Cholera Toxin; Epidermal Growth Factor; Mouth Mucosa; Epithelial Cells; Tretinoin; In Vitro Techniques; Epithelium; Culture Media; Pituitary Hormones; Blood; Transforming Growth Factor beta; Microscopy, Electron, Scanning; Animals; Humans; Keratins; Tetradecanoylphorbol Acetate; Calcium; Cattle; Cell Division

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.