A Human Epidermal Model can be Assayed Employing a Multiple Fluorescent Endpoint Assay and the Cytofluor 2300

Role of poly(ADP-ribose) polymerase (PARP) in DNA repair in sulfur mustard-exposed normal human epidermal keratinocytes (NHEK)†‡

“…Stock NHEK (passage 2) were cultured up to three passages in 5% CO 2 95% air at 37°C in a humid atmosphere 13 in KGM. Inducible PARP(Ϫ) human epidermal keratinocytes (HEK), previously described, 14 were cultured in a medium containing 75% KGM, 22.5% Dulbecco's minimum essential medium (DMEM) and 2.5% fetal calf serum in the same incubating conditions used for NHEK.…”

Sulfur mustard-stimulated proteases and their inhibitors in a cultured normal human epidermal keratinocytes model: A potential approach for anti-vesicant drug development

“…NHEK cultures were initiated in basal media from frozen stock (passage 2, P2) using 0.2 × 10 6 cells per 75 cm 2 plastic tissue culture flasks. Cells were grown at 37 °C in a humidified atmosphere of 95% air/5% CO 2 according to the method described by Rhoads et al [11] . When cells became approximately 80% confluent in the flasks, the cells were sub cultured to passage 3 (P3) to be used in experiments.…”

3.3. Eye Irritation

“…In the FL test, the corneal epithelium is mimicked in vitro by adherently growing epithelial cells, which form tight junctions and desmosomes in culture. The test is performed in a variety of formats, usually employing MDCK cells, although monolayers or multilayers of NHEK or human corneal cells have also been used (for example, 115,151,152). The cells are grown on inserts, so that, when confluent, the cell layer separates the medium into two compartments.…”