Transfection should i grow the cells in Media without antibiotic and FBS ?
It is my experience that some of the cell lines I have worked with are sensitive to having antibiotic (PennStrep) in the media during transfection by a reagent such as Lipofectamine. Once my cells are 70-80% confluent, I rinse off the antibiotic containing FBS-media and perform transfection in antibiotic-free FBS containing media. Depending on your cell type, excluding FBS during transfection (Serum starvation) might stress them out more... But again it depends on your cell type.
In my experience, FBS doesn't affect considerably the transfection efficiency in many cell lines but the use of antibiotics during transfection might be avoided since it interferes with lipofectamine and thus reduce considerably the transfection efficiency. Also transfecting cells under starvation condition for a long time may also trigger cell death(apoptosis). I recommend the use of an antibiotic free medium containg 5% FBS or optiMem.
Depending on the reagent mainly. Newer transfection reagents do not demand any medium change during transfection...even the cell lines succesfully transfected by the reagent are mentioned on the website of the supplier.
We use effectine and it works really well for a wide variety of human cell lines. Dont remove the FBS it has no effect and can hinder the process as cells may stall. We also keep in the pen/strep as it doesn't alter transfection efficiency.
Why not try the two conditions? Best way to learn how your cells behave. I'm always in favor of doing an extra condition, gives very usefull info. Good luck!
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