Hey everyone! i am trying to analyze the effect of a drug on mammalian cells. I know that I have to starve the cells before treatment but someone told me that I have to starve the cells again after treatment. According to my colleague, I need to starve the cells 2-3 hrs before extracting RNA and 8-12hrs before protein extracting. I dont really understand why i need to do this and according to several article, media starvation does induce the expression and activation of various genes. Can anybody help tell me if it is nessecary to starve the cells after drug treatment (Note:the drug is not a peptide). if it is necessary, would you be kind enough to give a reason. Thank you.
We dont starve the cells after treatment. And i did not find anybody talking about it even in the internet.
Lucy Evans,
We give serum starvation before giving drug treatment(6-24hr) for synchronization of total cell population(so that we get uniform results) , inducing differentiation (neuroblastoma) and creating stress. (Are you treating your drug in complete(10%FBS) media?) But we don't do post treatment starvation for RNA or for protein isolation. I guess post treatment induces the unwanted effects because cells in all treatments(for instance 1,10, 50, 100uM treatments) tries to regain their normal properties(I mean drug effectivity in concentration becomes nullified if you replace media with drug in to fresh media/serum free media).
Thank you all for your quick response. @ Chandra, I am treating the cells in 5% FBS.
You have not indicated what kind of drug, what kind of mammalian cells, and what effects you are looking for. However, experience and past work in the area of studying drugs ' effects on mammalian cells indicate that if your drug is forcing the cells to use an alternative pathway of doing something specific in the cells, there is no contraindication for the starving the cells since these cells are supposed to thrive on serum free medium. However, if you are getting a huge amount of cell killing or retarded growth, it is time to go with FBS or FCS.
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