Can anyone please suggest how many MCF 7 cells do I need in each well for optimized mtt assay? as you know, growth rate of mcf-7 cells is very high. is 5000 cells enough for each well ?
Hi! I've been working with MCF-7aro cells and, in terms of MTT assay, the cell density optimized by us is 2.5×10^4 cells/ml (2/3 days of treatment) and 1×10^4 cells/ml (6 days of treatment). Hope this helps!
Hi Aref,
I´ve been working with human oral fibroblasts. I used 10000 cells in a 96-well plate to perform the MTT test. The growth rate is not high. After 48h, I carried out the experiment by treating my cells with a product, during 30 min, 1h, 6h, 12h, 24h and 48h. This cell concentration was suitable for this cell type and duration of the experiment.
I think that if your experiment has a large duration and the growth rate of your cells is high, I woud try 5000 cells (maximum). But I think the most important is to work with a final cell confluence (in the moment of the MTT test) around 70-80%.
Best regards, Ana Celeste
Hi,
Ideally, you need to find the optimal number yourself. The growth of the cells may be different between labs. It will of course depends on the area of the wells and time of incubation before you start the MTT assay. Test different concentrations, e.g. in 96-well plates: 2000, 4000, 6000, 8000, 10000, 12000 cell/well and let them go untreated with the same time of your planned experiment. Then perform MTT and plot viability. You choose the highest concentration that match the linear growth. Stay away from concentrations were the MTT-signal flatten out, here the cells start to reach confluency.
Good luck!
- Badges
- Science method