Transfecting primary cells (imr90) - PROBLEM - (Sep/18/2006 )

Hi,
I am trying to transfect primary cell lines - in particular IMR90 - and have failed miserably.

So far the reagents I have used are:

1. Lipofectamine and Lipofectamine 2000 - Followed the instructions but they both lead to cell death! the few cells left behind were not transfected at all. Now I am going to do a titration with the reagents but dont have much hope there.

2. Fugene 6 - Cells were very happy but saw less than 2% transfection efficiency.

Does anybody have experience transfecting primary cell lines. Any suggestions???? I would very much appreciate some help to fix this issue!

IMR90 cells are really hard to transfect and i've switched for infections

Infection is a good solution, or maybe try with Amaxa nucleofection. I tried desperately to transfect FLS primary cells with a good yield (JetPEI, was too toxic, and I tried something else, but I can't remember the name right now), however Amaxa did the job greatly.

as I know for using Amaxa reagents you need a special apparatus from Amaxa which is not cheap; if classical transfection reagents fail, and there is no hope for improvement, one has to think about retro- or adenoviral infection; it is more complicated, & you need a special biosafety level lab but the chance to succeed will grow; another possibility is to select stably transfected clones even if at low transfection efficiency; upscale your transfection to get a good absolute number of transfected cells