Transfection - Cell death after transfection - (Jan/25/2005 )
I am having problem with transfection using Lipofectamine 2000 (Invitrogen). After transfection my cells dont survive even if I change the medium after 4 hrs of transfection. Any suggestion or advice is much more appreciated.
What are you transfecting, siRNA, antisense, plasmid? Both lipofectamine and DNA or RNA could be cytotoxical to cells. Have you done a mock transfection, ie. lipo only without vector? If yes, did you see the same with mock transfected cells? If yes, try decrease the amount of lipo used. Some cells are just very sensitive to transfection reagents.
If you have a very sensitive cell line, such as primary cells etc. you might want to give neophectin a go as it has a very hight transfection rate with very low toxicity. For more information, look at http://www.expresson.com/productsandservices/neophectin.html
Thanks for responses,
I am transfecting plasmid and am also doing mock transfection, will be looking the result at today.
Another good transfection reagent with much less cytotoxicity while maintaining high transfection efficiency is GenCarrier-1/2. We have being using it for a while without any problem on both primary and established cells.
Thanks Bryan for your suggestion
You may also reduce cell death by transfecting the cells when they are more dense (about 70%)