Good morning, I am trying to introduce an expression plasmid into THP-1 cells and have come across a couple of papers that describe electroporation techniques but on different machine than we have access to. However they don't give the actual parameters (they cite programme numbers instead). Does anybody have any suggestions on what might work or am I better to just go through a full optimisation? Alternatively, are there any other new transfection techniques out there that might work?
Personally, I always performed transfection of adherent cells. I had very good experience with Invitrogen reagents. At the moment I'm using this DMRIE-C transfection reagent (Catalog number: 10459-014) for adherent cells but it's given as "particularly effective for transfecting suspension cells".
Hello Cushla,
Please have a look to the attached file. It gives a protocol for plasmid transfection in THP-1 with Viromer RED. Expected efficiency is around 25% of transfected cells. To go up, I would suggest to transfect mRNA instead (if possible modified mRNA), you can then reach 50%.
Macrophages, monocytes, DCs... they are most of them quite "resistant" to plasmid transfection due to their self-defense mechanisms (e.g. AIM2/IL or CGAS/IF enzymatic cascades in response to foreign DNA). Transfecting mRNA is the best alternative.
Good luck and do not hesitate to ask me for further details.
Bests, Sandra
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