I am trying to transfect HPMEC using Lipofectamine 2000. I already tried transfecting for 6h using serum-free media (otimem) and the cell viability dramatically decreased, I reduced the transfection time to 3h using again serum-free media and the cells died, then I tried transfecting for 3h using complete media instead of serum-free and the cells also died. At this point I don't think decreasing the transfection time will do any good because of transfection efficiency. Does anyone know if Lipofectamine 2000 is just too toxic for endothelial cells? Is there a better transfection reagent that I can use? By the way, I am using around 50% confluent cells before transfection, should I increase the confluency? Could this really make a difference? Thanks.
Lipofectamine Plus.. is an improved reagent and used for difficult transfections..
Thank you, I actually tried Mirus LT1 and worked really well for this cell line, very low toxicity
Hi Virginia, what was your transfection efficiency for HPMEC using the Mirus reagent? Thanks, Domokos
Hi Domokos, I would say transfection efficiency was around 20%. See sample pics attached.
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