Submerse cultivation of NHEK and NHDF was performed at 37[degrees]C.
Investigations with NHEK and NHDF were carried out with cells from the 2nd to 6th passage.
NHEK isolated from human skin resectates and treated with geraniin and furosin showed increased keratin and involucrin expression compared to the untreated negative control as evaluated by semi-quantitative dot blotting of protein extracts with the respective mouse-anti-human-antibodies (Table 2).
Cell viability was measured in NHEK by neutral red dye uptake.
The levels of selected cytokines in cell culture supernatants from NHEK cultures before the addition of MTS or neutral red reagent (described above) were measured using the Luminex xMAP Protein Immunoassay according to the manufacturer's instructions (Biosource, Camarillo, CA).
Therefore, we evaluated the ability of CPE to induce cytotoxicity in NHEK.
Keratinocyte-derived cytokines mediate inflammatory responses in the skin, regardless of whether the stimulus is a result of allergen-specific or irritant effects, therefore, we evaluated cytokine profiles in NHEK following exposure to CPE (Figure 4).