Hi, what transfection reagent would you recommend for human primary endothelial cells? I have tried Lipofectamine 2000 on EA.hy926 cell line and on HUVEC, and it seems to be very toxic and the efficiency is below 1-2%.
Which is more easily transfected? dermal microvascular or pulmonary microvascular endothelial cells?
I used N-Ter transfection reagent from Sigma to transfect primary cultured HUVEC
It also depend on how much lipofectamine you add into the system. I have used lipofectamine 2000, LTX, and 3000 in sheep pulmonary artery endothelial cells and they seem O.K. to me most of the time. But, sometimes a specific primary cell cultures are just so sensitive and I have to dial down the amount after a few trials.
For HUVEC we use AMAXA, for Blood Outgrowth Endothelial cells thid doesn't seem to work. So what may work for one kind of EC's may not work for another kind. We also use lentiviral constructs often, that could also be an alternative way to obtain the required expression or knockdown.
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