Preparing a conditioned medium from cancer cells - (Nov/15/2012 )
I am working with HUVECs and the effects of VEGF on its growth and metabolism. I plan to use a conditioned medium from a cancer cell line. But I would like to know, if, in order to have enough VEGF or any other factor to activate angiogenesis in this medium is it sufficient that the conditioned medium be taken from growing the cancer cells in a normal manner? Is it not required to grow the cancer cells in a stressed or special (tumour micro) environment when these factors will be released actually, as in the real situation in human body?
If you are looking at VEGF it would be best if you could add this directly to the medium from a stock. Otherwise, how are you going to assay the levels in the conditioned medium between experiments to ensure that conditions are stable?
I don't know the answer to your actual question, other than to say that many cancer cell lines are happily secreting a bunch of different growth factors all the time, so it may not be necessary to stress them.
Well..this is what I am looking for..Cancer cells do secrete growth factors all times..but are they all angiogenic stimulants? If am looking for angiogenic stimulating factors in the conditioned medium (ok..not just VEGF..but may be others too) shouldn't I grow the cancer cells in special environment that induces them to produce such factors?
I do use a recombinant VEGF directly, but am also interested in random factors, as in the conditioned medium.
No, not all growth factors are angiogenic (as far as I know), so it is quite likely that there will be others in the medium. However, just because a growth factor is not angiogenic, doesn't mean that vascular cells won't respond to it....