transfection of lymphocytes - (May/26/2005 )

Hi,

I think it is quite difficult to transfect primary human lymphocytes with plasmids or proteins.
Has anybody experienced a way to do it efficiently and without significant cytotoxicity?

thank you

Primary lymphocytes are fragile and not easy to transfect.
We have some success using GenCarrier-2 on primary PBMCs with a NF-kB reporter. Efficiency is enough for luciferase assay with low cytotoxicity. DEAE-dextran is another way for the primary lymphocytes but with high cell loss and low efficiency in our hands compared to gencarrier-2.

using RNA, instead of DNA, by electroperation, you'll get over 90% transgene (GFP) expression with 80% viability, for both stimulated (with OKT3) or non-stimulated human PBLs.