Hi all,
I am trying to induce decidualization in primary human uterine fibroblasts.
The problem is that when i use charcoal stripped FBS instead of normal FBS, it causes a massive cell death.
Do you have any suggestion on how to prevent this?
Hello Ali Hedayatnia
Charcoal stripped FBS removes molecules necessary for cell growth and metabolism. It has reduced concentrations of steroid hormones, such as estradiol, progesterone, cortisol, testosterone, T3, and T4. The removal of essential proteins by charcoal stripping may cause changes in biological functions. So, when cells are in media containing 10% charcoal stripped FBS, trying adding supplements such as 3g/L glucose, 1mM sodium pyruvate, 1.5g/L sodium bicarbonate and 1% ITS (insulin, transferrin, sodium selenite).
When you are trying to induce decidualization in cells, you should use MPA and cAMP which are known inducers of decidualization.
Decidualization is induced through the concerted effect of cAMP and progesterone, or their respective analogs, 8-bromo-cyclic adenosine monophosphate (8-br-cAMP) and medroxyprogesterone acetate (MPA). Progesterone is necessary to induce as well as maintain the morphological and biochemical characteristics of decidualization in a long-term culture. On the other hand, cAMP’s main effect is to sensitize the cells to the action of progesterone. The combination of the two analogs induces faster morphological and biochemical changes than progesterone alone. All these changes are associated with an increased secretion of many proteins which are known to be markers of decidualization.
Best.
Dear Malcolm,
Thank you so much for your detailed answer.
Do you know any other supplements other than charcoal-stripped FBS which i can use instead of it for inducing decidualization?
Thank you so much.
Ali
Dear Ali,
Use serum free complete media deprived of hormones such as estrogen and progesterone as it may provide a better defined alternative to media supplemented with charcoal stripped-FBS.
Best.
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