1 reply to this topic

[eeri]

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

[miRNA man]

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.