I am trying to examine the cytotoxicity of an antioxidant agent on undifferentiated SH-SY5Y cells. In my previous try, all of the doses I tried gave results above 100% and all the wells were looking similar in color. I am not experienced in MTT assay, but I am thinking that I may change the doses. However, I am not sure how I should determine the new range. The doses I tried were 1 uM, 2 uM, 4 uM, 8 uM, 16 uM and 32 uM and I used 1 mM stock solution to dilute. I didn't change or skipped a step in the protocol but any ideas about the reason of failure is appreciated.
Dear Irem,
Most likely the matter is in the nature of the substance that you tested. If your substance is an antioxidant, then it will reduce oxidative stress in cells, which means it will stimulate their survival, then what you got (survival rate is more than 100%). If you want to get a cytotoxic effect, you need to use oxidants.
Best regards,
Alexander
Dear Irem,
I agree with the previous answer but also keep in mind that the dose makes the poison. So if you want to see any cytotoxicity, maybe you can increase even more the dose range. Also, you can look at the literature on that antioxidant agent and see if something has been done related to your work.
Best,
Ana J.
An antioxidant agent is not necessarily a cytotoxic agent. An agent can have antioxidant activity but not cytotoxic activity. I agree with both the previous recommendations.
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