5.6. Cytotoxicity assays using human keratinocytes

Human keratinocytes from histologically normal skin (HaCaT cells; Fisher Cat # T0020001) were maintained in DMEM High Glucose media with 4 mM L-Glutamine (NyClone) and 100 U/mL penicillin, 100 μg/mL streptomycin (pen/strep) (Life Technologies), and 10% (v/v) inactivated fetal bovine serum (FBS) (NyClone). Cells were grown in 5% CO₂ at 37°C and were detached from culturing dish at 80% confluency using 0.025% trypsin-EDTA (NyClone) treatment. Peptide influence on the cell viability was estimated by using the MTS assay according to manufacturer’s protocol (MTS, CellTiter96 AQ One Solution Cell Proliferation Assay, Promega).