I want to investigate the fish oil cytotoxicity using MTT Assay
Should I extract the active ingredients from the fish oil first then apply the extraction on the cell
OR
I can add the fish oil directly on the cell with media
Hello Hazem Choukaife
You should extract the active ingredients from fish oil first and then test the extract on cells for cytotoxicity. The active ingredients are substances with biological activity that would give a much clearer idea of potency than just using fish oil directly on cells.
So, to ensure the effectiveness of fish oil, extracting the active ingredients and checking for cytotoxicity would be appropriate.
Best.
Malcolm Nobre many thanks for your answer
Yes because if I want to add the fish oil directly, it will not be mixed with media
I would say to test the crude oil first for its cytotoxic effect and proceed to the isolation of the active components only after confirming their presence with the MTT Assay. If the crude oil does not show potent cytotoxicity, isolation would be unnecessary. Directly isolating actives may cause you to waste time and resources if none of them shows good results. Also, hydrophobic substances will not mix with your media therefore you will have to find a suitable drug delivery method as well.
You can emulsify the fish oil with your media, your media likely has FBS in it, and the albumin in the FBS has the capacity to carry fatty acids and lipids. Do not use sonication to aid in the emulsification process under aerobic conditions (you would need to purge with nitrogen to remove the oxygen first). This is because sonication under such conditions will result in the formation of lipid hydroperoxides and other ROS species. It should be noted that free heme or hemoglobin (and possibly other heme proteins) in the media can catalyze lipid peroxidation. See below paper.
Article Hemoglobin Is Not a Biological Fenton Reagent
Good luck.
F. Nghakliana So you prefer using crude oil as reference,
many thanks
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