I have made a cytotoxicity assay using MTT at 20K cells/well in a 96 wells plate.
If I do a MTT assay at 50K cells/well and at 10K cells/well, will i found the same effect ?
I tried to find some previous papers about that but did not found anything relevant.
Definitely the number of cells can vary the cytotoxicity effects on the cells. The amount of time you culture the cells will also have the effect. In general, I plate 10k-20k cells/well in a 96 well plate in order to conduct MTT cytotoxicity assays. This allows the cells to not be fully confluent, allow for attachment for overnight, and then proceed with the experiment as necessary with appropriate time points.
So, you have to optimize the assay condition regarding cells numbers, cell attachment timing, cell doubling time. You are highly advised to do a cell number titration to see the cell growth rate.
Hope that Helps.
@Masuma Akter okay thanks. But if i conduct a study at 100k cells/well, how can i measure cytotox with MTT ? Should i use another method ?
If you seed very high number of cells e.g. 100K cells, they will definitely be in exponential phase within 10 hours from seeding (assume normal condition of cells; if disease that can vary). Based on your cells grow very fast or grow very slow, then seeding number can be varied.
In case of high number of cells e.g. 100K cells seeding condition, you can do MTT assay
Hope this helps.
Yeah. it's vero cells, 24h cycle. So instead of 48h hours to read mtt i should wait 24h to see something maybe ?
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