3 replies to this topic

[Monik]

Hi all!

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

[expresson_help]

NeoPhectin is extremely low toxicity compared with a lot of other transfection reagents - this might help you. More information at http://www.expresson...neophectin.html

David
Expresson

Edited by expresson_help, 18 January 2005 - 05:12 AM.

[thpproblem]

Hi,
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...

[thpproblem]

once again...hi!

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...

cu