Materials
- Cell lines: A549 cells (American Type Culture Collection)
- Cell culture growth media (Dulbecco’s Modified Eagle’s Medium (DMEM) supplemented with 10% Fetal bovine serum and penicillin/streptomycin)
- Other cell culture components: Phosphate Buffered Saline (PBS), Trypsin-EDTA (0.25% trypsin, 1mM EDTA)
- 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT)
- Silverlab Healthcare sample (IM/Q610/419-01A-18).
- Luminometer
Laboratory equipment and consumables
- CO 2 incubator
- Biosafety cabinet
- Bench top centrifuge with an adaptor for 96 well plates
- Water bath
- Flat bottom 96 well plates
- Flat bottom 96 well black plates
- T75 tissue culture flasks
Method
Cytotoxicity assay for IM/Q610/419-01A-18 on A549 lung cells
The cytotoxicity assay for the IM/Q610/419-01A-18 sample was performed by first seeding 10 000 cells/well/100 µl of A549 cells in a 96 well plate for 24 hours at 37°C, 5 % CO and 95 % humidity. After 24 hours, a 2-fold serial dilution of the sample was performed and 100 µl was transferred to the plate containing cells, except in the cell control wells.
The cells were then incubated for 72 hours. Following the 72 hours incubation media was removed and replaced with 25 µl MTT reagent (5 mg/ml) (Sigma- Aldrich, St. Louis, MO). This was followed by 3 hours incubation at 37°C to allow the formazan product to form from viable cells. After incubation, the MTT was removed and 100 µl DMSO was added and incubated for 15 minutes at room temperature. Afterwards the absorbance was read at a wavelength of 620 nm using the Tecan Infinite F500 luminometer.
