I want to do an experiment where I take conditioned media from one plate of cells and and add it to a plate of cells with a gene knockout and see if there are changes in phenotype. Can someone run through a protocol on how to do this. Do I add serum in the plate while I am conditioning the media but no serum to the plate of cells for which I will be adding the conditioned media to or no serum at all? These are lung tumor cells which normally take RPMI with 10% FBS.
Conditioned medium is likely to include some kinds of cytokines or growth factors which render the phenotype of the recipient cells, so that you do better use RPMI without 10% FBS. After all, serum is rich in not only nutrition and amino acids but also growth factors and cytokines!!! The presence of the FBS in the medium of recipient cells may mask the real influence of the conditioned medium.
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