I am making a toxicity test with ruthenium in gallbladder cancer cells, some references use time of incubation with drug of 24 and 48 hours. But ¿what is the basis for this? How I can determine the incubation time should I use?
thanks for your attention
Hi Hernan,
If you are testing a compound/drug for the first time in vitro, the best approach would be to do a preliminary screen to determine the changes in morphology over 12,24,48 and 72 hrs. This will give you an idea of where cell death happens and possibly if your control cells survive the longer treatment times or not. Here you have to test at 3 broad concentrations ( or better determine it based on previous literature of any).
Based on what you observe here, you can establish a suitable incubation time. In my opinion 24 hrs would be a better strategy to report. But a lot of them do report at 48 hrs. It depends on what you observe and what you would like to report. In your case, if it is only toxicity that you want to find out, then if you see anything within the first 24hrs itself, then the compound has a higher propensity to be toxic to your cells
I agree with what Harini wrote. Actually the time it takes for a drug to exert its action is a very interesting aspect of pharmacodynamic, so it is very likely that you will have a different IC50 for different exposure times. Generally the concentration required will be higher for shorter exposure times: you can also test a short exposure (1 to 24 hours) followed by a wash-out phase (you change the medium and do not put any drug). Int hisway you study a sort of hit-and-run mechanism.
This is the most interesting part of studying new agents in vitro!!!
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