The culture of human buccal and vaginal epithelial cells for permeability studies

The objective of the present study was to develop a single improved technique to culture human vaginal and buccal epithelial cells, whereby the cultured cells can be used in drug permeability studies. Cells were obtained from healthy human vaginal and buccal mucosa following vaginal hysterectomies and various oral surgical procedures. Tissue obtained was washed extensively in phosphate buffered saline (PBS, pH 7.3) containing antibiotics and amphotericin-B. Tissue specimens were cut into small pieces and plated out in 24-well plates. After drying, the full medium was added. Cell growth occurred within 4-6 days from primary explants and confluency was reached within 2-3 weeks. Primary explants yielded epithelial cells with minimal fibroblast contamination. After trypsinization, cells were seeded into collagen-coated wells and onto Transwell membranes. Trypsinized cells grew best on collagen-coated surfaces yielding more than one layer. The average steady state flux for the collagen-coated membranes (ccm's) containing either buccal or vaginal epithelial layers towards water was 6-10 times lower than that found for the cell-free ccm's. Fluxes for cultured cells on ccm's were 3x higher than those obtained for intact buccal and vaginal mucosa. Growth and permeability to water of the vaginal and buccal epithelial cells were comparable, confirming the similarity of these two tissues and the suitability of using the former as a model for the latter in permeability to tritiated water.