Do you think the background level of E2 in culture medium supplied with FBS 10% could effect on the E2 measure after treatment granulosa cells with factor? As my opinion, we had the base levels of E2 between group, then it is not effected to the result. Please advise me with this thought? Thank you so much.
If you have concerned, you can make serum steroid-free by treating with charcoal. One of the procedure is from Sigma (attached). Think about it. E2 level will vary between batches of Fetal serum.
Dear Dr. Subhash C. Juneja thank you so much for sending me this procedure. I worry when we use charcoal, not only of steroid but other growth factors need for cell culture also lost. I understand that E2 will differ between lot to lot, if we use the same lot for repeat experiment, could we believe in the result. I wonder this question because of this medium, I think it concern when you treatment cell with steroid. In case of observing the steroidogenesis effect of treatment factors, this may not a problems. I try to find about the research concern about this problem, however, I could not find any paper about this issue. Thank you so much for answer my question.
E2 is E2; and the background level will affect your experiments unless you have added considerably more E2 than what was already there. The E2 target level or a range for the desired E2 concentration should probably be a part of your experimental protocol. Your title suggests (to me) that you can measure the E2 concentration in the serum you use. There may be kits for this that you could find. If you can not measure the level, the serum supplier may routinely be able to provide this information to you. You might start by asking the serum supplier for a recent batch and then ask for serum from the same batch lot number if you have to order serum again. I suggest that you always check the serum lot number on the product you have purchased and record the lot number in your records for future reference. You might also keep track of your storage conditions, such as refrigerator and freezer temperatures.
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