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cells dying: MCF-7: siRNA transfection + estrogen treatment


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#1 eeri

eeri

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Posted 27 August 2009 - 11:57 PM

Hi,
I am struggling with a serious problem: after siRNA transfection MCF-7 cells die.

Since I want to treat the cells with estrogen later on, I usually grow them for 2-3 days in phenol-free medium containing 10% charcoal-stripped FCS. Then I transfect the cells with siRNA using lipofecatmine RNAiMax and leave the transfection mix on the cells. After 1 day cells do not look great.

however, after siRNA transfection with normal medium, normal FCS cells are OK, while after siRNA transfection in the steroid-free medium cells die. Has anybody an idea what can be improved? which E2 concentration do you usually use, how long do you treat the cells with E2 then?

Many thanks,
eeri




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