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.