transfection on THP-1 cells - a BIG problem (Nov/25/2004 )
I'm trying to obtain a silencing in THP-1 cell using commercial siRNA after which I have to stimulate my cell to induce cytokine production.
Before transfection, I have the need to differentiate THP-1 (they became adherent) to induce receptor on cell surface and only after this passage I can transfect the cells.
I have a lot of problem to find a reagent for transfection that minimally influence cell viability (> 50% cell death when all seems to go well!!!!!!!!) and I think that the preliminar differentiation step is critical for cell healthy, because I always obtain great differences in transfection efficiency.
Is there someone that can suggest me how overcame my difficulties?
Thank you all
NeoPhectin is extremely low toxicity compared with a lot of other transfection reagents - this might help you. More information at http://www.expresson.com/productsandservices/neophectin.html
I´ve the same problem...
do you find a solution for transfecting thp-1 cells (after differentiated to macrophages)???
perhaps you can take a look at amaxa.com...they seem to transfect non-dividing cells...
worst case approach:
perhaps you can tranfect the thp-1 cells before you treat them for differentation
i guess they need about 48h for differentation, maybe the transient expression of your siRNA will be > 48h...