I am going to transfect plasmid in U2OS cells using FuGENE HD/Viafect transfection reagent plus Opti-MEM as a medium. Does anybody have experience with this particular cell line?
Could you please suggest effective cell density/ reagent:DNA ratio etc for 4 well plates.
Well, I obtain good results using Lipofectamine2000, I don't know if they work in the same way...any way, I use cells at 80% of confluence and for the 4 wells I plate 25000 cells and 2 days after I do the transfection. The DNA ratio is something that depends on your experiment, for example I have good results by adding 75ng for one plasmid and 250ng for other.
U2OS cells are quite easy to transfect! Don't you worry :)
Hi Debora and Zulfikar, i recently started transfecting U2OS cells siRNA against my GOI. I am observing bright vacuole cytosolic spheres after transfection. Interestingly these are more abundant in my sample (siRNA for GOI) as compare to control siRNA. I checked with other cell lines, but i could not find. Did you find any kind of vacuoles after transfection with effectene? How cells appear after transfection (with respect ur transfection reagent)?
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