Hi, I'm following a protocol to produce Rspo1 conditioned media by using 293T cells. I was wondering why it is recommended to prepare the conditioned media by treating the cells with AdDMEM f12;1% glutamax and 1% hepes without using FBS. The FBS is instead present in in the complete media they recommend to use for cultivating these cells.
I am also producing Wnt3a conditioned media and in that case the media I use to cultivate the cells and to prepare the conditioned media is the same and there is 10% FBS in it.
Do you have an idea? It is the same if I add FBS when preparing Rspo1 conditioned media?
Thank you in advance
FBS is used as a crude source of various growth factors. With that protocol they are wanting the cells to condition the media by metabolizing but do not want them to proliferate too much which would deplete the media for your actual experiment.
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