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[SF_HK]

I'm doing MTT to check the effect of a drug on the proliferation of a cancer cell line. I've seeded about 2,000 cells / 100ul in each well.  The medium won't be enough to sustain the cells for 5 days. In such a case, do I replace the medium (with the drug) with fresh medium containg the drug, after about 3 days?

Thank you.

[bob1]

That should be OK, though this would be equivalent to giving two doses of the drug.  Do you know much about the physiology/pharmacology of the drug and how it is broken down/metabolized and the effects of the metabolites?