Preparation of Conditioned Medium - (Nov/06/2011 )
I'm testing conditioned media of different tumor cells to assess the ability to cause proliferation. However, I did not do any normalization between this mediums. I wonder what would be the best method to normalize these mediums to eliminate interference by different concentrations of factors among them. And how should I prepare them? Many protocols that I read say that medium should be centrifuged and filter. But I do not see why centrifuge and filter. Centrifugation would not remove all debris already?
Thanks a lot!
Centrifugation might not remove all the particles, depending on the RCF used. Filtration will also remove any potential bacteria that you may accidentally introduced.
If the conditioned media are all from the same cell line in the same medium, just combine them all in a large container such as a T-150 flask, then aliquot and proceed as you would normally.
If they are in different media or from different cell lines, I don't think there will be any real way you can normalize between the different batches.