Skanit sotware 3

To assess cellular toxicity, MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) tests were carried out. For the experiments, cells were seeded (5 × 10³ cells/well) in 96-well plates in standard culturing medium for 72 h at 37 °C, thus allowing cell adhesion and confluence before changing to serum-free media (SFM) for 24 h. After 24 h, SFM was removed and replaced with fresh SFM containing liposomes at different phospholipid concentrations. Untreated cells with only SFM at every plate were used as controls. Following incubation for 24 and 48 h, cell proliferation was assessed by MTT according to the manufacturer’s instructions (Roche Life Science, Indianapolis, Indiana, USA). Briefly, 10 μL of MTT reagent 1 (MTT labeling reagent) was added to each well and incubated for 4 h at 37 °C before adding 100 μL of MTT reagent 2 (solubilization solution, 10% SDS in 0.01 M HCl) and incubating at 37 °C before reading plates. The samples were measured on a Multiskan GO plate reader (test wavelength 595 nm, reference wavelength 660 nm) using the SkanIt Sotware 3.2 (both Thermo Fisher Scientific, Waltham, MA, USA).