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MCF-7 cells dying after siRNA transfection + estrogen


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#1 eeri

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Posted 28 August 2009 - 12:02 AM

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?

Many thanks,
eeri

#2 miRNA man

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Posted 28 August 2009 - 01:48 PM

What happens if you culture the cells in the stripped medium and don't transfect them? Do they also die? Maybe you should also look into a different transfection reagent, or using less of your current lipofectamine, since transfection stresses cells.




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