transfection of suspension cells - how to improve the transfection efficiency? (Aug/01/2006 )

hi,

I find that the transfection of suspension cells is much difficult than that of adherent cells, and transfection efficiency is very low.

Could you give me some suggestions? Thank you very much!

It is the origin of the cells that matter. Suspension cultured 293 and CHO are very easy to transfect. On the other hand, most cells derived from white cells are typically hard to transfect. 0.1-10% if you are lucky. You have better luck with electroporation.

I transfect 293s with lipofectamine 2000 in suspension.

i used to electroporate BHK and got good transfection efficiency as well.

It depends on your cell line, I've had varying transfection efficiencies on different suspension T-cell lines, ranging from about 40% 'till less than 2% (determined by FACS with EGFP reporter gene). They were transfected with the same DNA, I made sure they were exponentially growing and all that.

I'd say you better try to find the right transfection reagent or try to transfect with different ratio's of transfection reagent to DNA.

Good luck!