Could someone that work with human mo-DCs transfection with siRNAs recommend a best method to do this? I generally have seen works using lipofectamine 2000 (Invitrogen), eletroporation (Amaxa/bio rad) or lentivirus. I need a method with high efficiency that obtain great number of viable cells to infection experiments and pathogen-host analysis
Hi Amanda
I have used FuGENE® HD from Promega to transfect Phoenix_Ampho cells and it worked pretty well for me. It was gentle on the cells and it gave a high transfection effeciency. In my opinion, you can use FuGENE® HD to get a high transfection effeciency with the DC cells.
Finally, I have attached a recent article in which the authors successfully used FuGENE® HD to transfect a number of different cells including the DC cells.
Transfection of DCs is tedious, because of the high mortality of the cells using reagents such as Amaxa. We have been succesful with the use of lentiviruses to knock-down genes from murine BM-DCs. The lenti infection allows to work with a decent number of live, immature cells, for applications such as antigen presentation, microscopy, phagosome maturation, among others.
You can check recent papers from our lab:
- Innate Immune Signals Induce Anterograde Endosome Transport Promoting MHC Class I Cross-Presentation.
- Impact of the TAP-like transporter in antigen presentation and phagosome maturation.
our Viromer BLUE and Viromer GREEN have shown great efficiency for DCs and other immune cells. Please, check table below for list of citations (with links to find optimized protocols).
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