MTT assay

Effects of DHA on cell proliferation was determined using an MTT assay. The concentration of the cells was adjusted to 4×10⁶ cells/ml, and 190 µl cell suspension was added into each well of the 96-well plate. Plates were incubated for a period of 24 h using a 5% CO₂ incubator at 37°C. Various concentrations of DHA (1, 3, 10, 30 and 100 µmol/l) were added to the plates and incubation was continued for 12, 24 and 48 h. The addition of 20 µl MTT solution (5 mg/ml) to each well of the plate was conducted at 12, 24 and 48 h. Further incubation of the plates was carried out for 4 h, followed by decantation of supernatant and the addition of 150 µl dimethyl sulfoxide to every well. Measurement of the absorbance for each well was performed in triplicate at a wavelength of 490 nm to determine the cell viability.