Hi, Alireza,

FBS contains proteins and other molecules that may interfere with the action of drugs on cancer cells. For example, certain proteins in FBS could bind to drugs, reducing their availability to interact with their molecular targets within the cancer cells. This interference can affect the efficacy of drug treatments and may lead to inaccurate assessment of drug potency or sensitivity. To mitigate the effects of FBS on drug treatment in cancer cells, researchers often explore alternative culture conditions, such as serum-free media or the use of defined serum substitutes. These alternative approaches allow for better control over the cellular environment and can reduce variability in drug response between experiments.

Hello Alireza Ghorbanpour Khaljiri

Besides having anti-cancer properties, Baicalein is also a potent antioxidant and has anti-inflammatory properties. Baicalein is a flavonoid found in the chinese herb, Scutellaria baicalensis. It is an unstable flavonoid with a pyrogallol substitution in ring B. It shows instability in DMEM medium at 37 deg. C.

You may want to refer to the articles attached below for more information.

https://dora.dmu.ac.uk/server/api/core/bitstreams/dc38f816-8224-4fa8-beeb-e07591706d6b/content

https://www.nature.com/articles/s41467-021-23751-3#:~:text=Notably%2C%20the%20pyrogallol%20group%20of,state%20of%20the%20proteolytic%20reaction.

Best.

A few points to consider Baicalein can auto oxidize generating species that can cause oxidative stress (depending on levels the levels of ROS generated). Moreover, several of the species generated can reduce MTT directly.

Article Baicalein, as a Prooxidant, Triggers Mitochondrial Apoptosis...

All of these factors can complicate these experiments.

You can also try to add a color control to your assay. So you do a color control for each concentration of your compound and subtract the effects from your test samples. A color control is your drug (you do it for each concentration) and your media only, no cells. This should tell the real activity of the compound on cells that is if there is no interaction between the various components of your media and the compound.