Filter‐grown TR146 cells as an in vitro model of human buccal epithelial permeability

Abstract

Use of a cell culture model of a specific epithelium requires documentation of its differentiation. This study reports permeability of mannitol concurrent with a profile of differentiation markers of filter-grown TR146 cells, a cell line originating from a human buccal carcinoma, cultivated submerged or at the air-liquid interface for 23 to 31 d. A multilayered squamous epithelial-like tissue was found. The maximal permeability barrier and the most distinct stratification were obtained at day 23, when cultured submerged (apparent permeability coefficient 4.08 +/- 0.15 (x 10(-6)) cm/s; transepithelial electrical resistance 102 +/- 5 omega x cm2). The profile of differentiation markers demonstrated similarities to normal human buccal epithelium by expression of K4, K10, K13, K16, and K19 keratins, plasma membrane-associated transglutaminase, involucrin, and epidermal growth factor receptor. Uniform expression of K5, K8 and K18 was consistent with the carcinogenic origin of TR146 cells. Identical profiles of differentiation markers were obtained irrespective of method or time of culture. Karyotyping proved the human origin of TR146 cells. Three different passages had near triploid (3n+-) chromosome compliments and consistent occurrence of four marker chromosomes [mar4, mar5, mar9, and add(5)(p)], while differences between them defined them as subclones. The results indicate that a submerged filter-grown TR146 cell culture at day 23 of culture has the potential to model the human buccal epithelial barrier for permeation of drugs.

Cytotoxic and DNA-damaging properties of glyphosate and Roundup in human-derived buccal epithelial cells

“…The medium was changed every 2-3 days; when the cultures reached confluency, the cells were washed with Dulbecco's Phosphate Buffered Saline (DPBS, Lonza, Verviers, Belgium), detached with TrypLE Express (Gibco, Invitrogen, Carlsbad, CA, USA), centrifuged, and subcultured. TR146 cells express ultrastructural characteristics of normal human buccal epithelial cells, e.g., intermediate filaments, microvilli-like processes, and lack of complete keratinization (Jacobsen et al 1999).…”

“…Primary aim of the present study was to find out whether exposure to G and R causes adverse effects in the humanderived buccal epithelial cell line TR146 (Jacobsen et al 1999). In cytotoxicity experiments, four different endpoints were used, which reflect different modes of action.…”

Oral uptake of nanoparticles: human relevance and the role of in vitro systems

“…The best-characterized in vitro cell line that recapitulates the stratified squamous epithelium in the oral cavity are TR 146 cells, which are isolated from a lymph node metastasis of a buccal squamous cell carcinoma (Boukamp et al 1985;Rupniak et al 1985). TR 146 cells can be cultured on transwells, differentiate into a non-keratinized epithelium, and reach their highest transepithelial electrical resistance (TEER) value after 28-30 days (Jacobsen et al 1999;Nielsen and Rassing 2000;Nielsen et al 1999). They form 4-8 epithelial cell layers corresponding to a thickness of approximately 60 µm.…”

Evaluation of tissue engineered models of the oral mucosa to investigate oral candidiasis

“…However, the oral mucosa is a complex multi-layered, multi-cellular tissue consisting of increasingly differentiated epithelial cells and so monolayer cultures are unlikely to be representative. Recently, multi-layered organotypic three-dimensional in vitro culture systems have been developed to mimic the oral mucosa [17][18][19][20]. The reconstituted human oral epithelial (RHOE) model, which is based upon the TR146 buccal carcinoma cell line cultured on a porous polycarbonate insert, has been widely used in C. albicans infection studies [21][22][23][24][25].…”