2.11. Cytotoxicity of fusion peptides
The cytotoxicity of fusion peptides was assessed using CytoTox 96® non-radioactive cytotoxicity assay according to the manufacturer’s protocol. Human dermal fibroblasts neonatal (HDFn) cells were cultured in DMEM medium supplemented with 10% FBS and 1% penicillin-streptomycin at 37°C in 5% CO2 humidified incubator (Esco Micro Pte. Ltd., Changi, Singapore). HDFn cells were seeded on a 96-well plate at a density of 8.0×103cells per well. After overnight incubation, serially-diluted fusion peptides were added to cells at concentrations of 1, 0.5, 0.25, 0.125 and 0.0625 mg/mL. After incubation for 5 hours, 50 μL aliquot of each well was transferred to each empty well of a 96-well plate. Then, 50 μL of CytoTox 96® reagent was added to each well. Cells were incubated for 30 minutes at room temperature under light-free conditions. After adding 50 μL of stop solution, absorbance was measured at 490 nm using a microplate reader.