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transfection on THP-1 cells


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#1 Monik

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Posted 25 November 2004 - 01:47 AM

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

#2 expresson_help

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Posted 18 January 2005 - 05:11 AM

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


#3 thpproblem

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Posted 25 January 2005 - 06:10 AM

Hi,
Ive 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...

#4 thpproblem

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Posted 07 February 2005 - 11:55 AM

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




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