Hello all,
I am working with hormone responsive cancer cells (MCF7, ER positive) and wondering whether I need to use stripped serum or not .
In literature, some people either strip their serum to deplete it from undefined levels of hormones like estrogen and then add it back but sometimes they do not stripp serum indicating that fetal serum does not contain hormones in contrast to bovine serum .
Does anyone have ever done some analysis or could tell me where this has been demonstrated or what I could refer to once I have to prove it?!
Thanks for any suggestion
Generally, fetal bovine serum (FBS) is rich in a diverse range of hormones/ growth factors. This quality is desirable and explains why demand and cost is so high for FBS. Adult animal serum will generally contain fewer of the desirable growth factors, but will contain hormones. Serum contents are highly variable in all cases. If you wish to know the contents of your serum you can either test it yourself, or buy pre-tested serum from a reliable vendor. If you contact your vendor, and give them the lot number of your serum, they will be generally be able to provide this information for you. It would probably be of little use to have the test results from someone else's serum.
Hello Vanessa,
Actually, FBS does contain low levels of estrogens. You also need to know that phenol red has some estrogenic activity. I used to work regularly with MCF-7 cells before. We were using phenol red-containing medium with regular FBS for the maintenance of the cell line. When we were planning an experiment involving estrogen receptor signaling, we were pre-starving a subculture in phenol red-free medium containing charcoal-dextran stripped FBS for one passage before seeding the cells for the experiment in the same estrogen-free medium. That ensures that you get the maximal sensitivity on estrogen induced signaling pathways detection when you compare your estrogen treated sample to your untreated control.
Regards.
Thanks Mr Hayakawa,
unfortunately your answer is not really helpful for me as someone who is not familiar with the method, thanks anyway.
@Martine Bail :
So I use white Medium and informed about stripping serum, but wondered if I could avoid it ....
I let the cells that were maintained in red meidum with normal FBS now 48h in white Medium with stripped serum.
Their cytoplasm enlarged pretty much and they look unhealty...... did you also notice that ?
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