Cells were
trypsinized with
trypsin 0.25%, counted by
trypan blue (Biosera, France) and suspended at a concentration of 3 × 10
4 cell/ml. 100 μl of this suspension was added to each well of a 96-well plate. Cells were left overnight to attach to the plate. After 24 h, cells were inoculated, in triplicates, with quadruple-serially diluted concentrations of the algal extracts ranged from 6.25 μg/ml to 400 μg/ml. Negative controls for each extract concentrate were normal media containing the corresponding
DMSO concentration. Doxorubicin was used at double-serially diluted concentrations between 0.9 × 10
−2 μg/ml to 0.58 μg/ml. After 72 h of treatment, the medium was removed, and cell viability was measured by MTT assay. Briefly, 100 μl of MTT (0.5 mg/ml) in RPMI-1640 without phenol red was added to each well in the dark, and the plates were placed in the incubator for 4 h. Next, the supernatant was removed carefully, and 150 μl of
DMSO (Merck, Germany) was added to each well. Plates were shacked for 15 min and were then read at 570 nm using a microplate reader. All the experiments were repeated three times. Cell cytotoxicity of each extract was calculated using the following equation:
%cytotoxicity = 100 − (ABS
test/ABS
control × 100%).
Where ABS
test is the average absorbance of cells treated with algal extracts, ABS
control is the average absorbance of corresponding
DMSO control.
Erfani N., Nazemosadat Z, & Moein M. (2015). Cytotoxic activity of ten algae from the Persian Gulf and Oman Sea on human breast cancer cell lines; MDA-MB-231, MCF-7, and T-47D. Pharmacognosy Research, 7(2), 133-137.
