ABSTRACT
This chapter presents some current in vitro cell or tissue culture methods that have been proposed for in vitro evaluation of dermal phototoxic compounds. In most cases of phototoxicity, endogenous or exogenous chemical absorbs light and transfers the energy to, or reacts in the excited state with, cellular components. In experiments, human keratinocyte preparations were made from electrokeratomed strips of skin 0.5 mm thick. Lipoperoxide measurements were then performed using a reconstructed human epidermis that was exposed to a pure UVA waveband. The strips were incubated in antibiotic solutions, then placed overnight at 4°C in 0.2% trypsin-EDTA in calcium-, magnesium-free phosphate buffered saline. Human keratinocytes seeded on this dermal substitute were grown submerged for 3 days in a medium consisting with Dulbecco’s modified Eagle’s medium/Ham’s Fl2 supplemented with hormones, growth factors, and fetal calf serum. The culture was then exposed at the air-liquid interface for 10 days in order to obtain a fully differentiated epithelium.
