We have a transformed mouse macrophage line (PMJ2) and we are having trouble transfecting it. We used Lipofectamine but very poor efficiency. We don't have electroporation equipment available so we're looking for something else.
It looks great but we don't have the Amaxa or any other electroporation equipment.
I've used Hiperfect (Qiagen) fot transfecting pma-differentiated macrophage-like human THP-1 cells (efficiency around 70%) and primary human monocyte-derived macrophages (efficiency around 90%). The primary macrophages exhibit very little cell death during transfection desite the fact that rather high amount of Hiperfect needs to be applied (8 ul per 12-well). The THP-1 cells were more sensitive.
Thanks Kristiina, I haven't heard of that reagent. Did you follow a protocol that came with Hiperfect or did you have to adjust from there?
I notice I left out the crucial fact that it was siRNA that I was transfecting to macrophages with Hiperfect; I started with the manufacturer protocol for THP-1 cells which was a good starting point, yet for these cells I had to modify seeding density & pma-differentiation stage before it worked. Hiperfect is recommended for transfection of eukaryotic cells with siRNA and miRNA.
Another reagent that worked nicely for primary human macrophage transfection was the Lipofectamine LTX PLUS, though the application was a bit different, I transfected poly(dA:dT) i.e. repetitive synthetic dsDNA for immune stimulation.
Thanks for the clarification. The use of Lipofectamine LTX PLUS for delivering dsDNA is interesting. The TLR for that is in the nucleus or in the cytoplasm?
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