I'm assessing the effect of different doses of various drugs on Breast cancer cell lines.
All drugs were dissolved in DMSO.
Using MTT assay, the impact of some drugs on proliferation was evident. I then assessed whether the effect is cytotoxic or anti-proliferative using caspase-activity kit. The result indicated no cytotoxicity. Yet, I wonder if the solvent of these drugs, i.e. DMSO, contributed in any way to the results I've.
My question therefore is: how to ensure that the effect on proliferation seen by MTT is that of the drug not DMSO?
There are quite a few other solvents, or solubilizers you could use for example. DMF, DMAc, Acetonitrile, Acetone, β-Cyclodextrin (β-CD)
Hello Mariam Ahmed Galal
In MTT assay, have you included a control with DMSO in culture media, but without the drug? Use the same concentration of DMSO that is present in the treatment group. This will indicate whether DMSO alone has any effect on proliferation.
Best.
DMSO has toxic effect on cells so limit the concentration as much as possible. Usually less than 5 % DMSO is considered to be safe enough for cell.
Philip G Penketh Thanks for the suggestions. so these options are not known to be toxic to cells?
@Malcolm Nobre I've not really done this yet but now that u mentioned it, I'll give it a try. Really appreciate ur help. Thank you very much
@F.Nghakliana most effects were seen at below the 5% concentration so that's kind of a relief till MTT proves it rit. Thank you so much
Well everything is essentially toxic at some concentration, even if it is just outside of a normal concentration ranges. However, by measuring the concentration required to solubilize the MTT, the toxicity of the vehicle can be minimized by using the lowest concentration you require. Furthermore, different vehicles will have different toxicities. You will likely have to do a few experiments to determine which vehicle exhibits an insignificant effect on cell growth at the concentration required. Good luck.
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