I differentiated THP-1 cells to macrophages using PMA(100ng/ml) and then used lentivirus to deliver shRNA to knockdown my target genes. Unfortunately, it was too difficult to be transfected efficiently. I would like to know if once they are PMA-treated, how can the cells be efficiently transfected with lentivirus? Is siRNA easier for macrophages to be transfected? Is there any good reagent for siRNA transfection?
For unstimulated and differentiated THP-1 cells the Nucleofection (amaxa / Lonza Nucleofector) does work excellent. The same transfection protocol can be used for DNA but also siRNA transfection. As customer references I would like to mention the Lorkowski paper, also video publication of JoVe:
Efficient non-viral transfection of THP-1 cells: Article Efficient non-viral transfection of THP-1 cells
I would like to note that one can also infect undifferentiated THP-1 cells and select the infected cells using a drug (e.g., puromycin) and then differentiate the cells using PMA.
I used Viromer Green for siRNA transfection of THP-1 Cells. I would transfect cells before differentiate them. So I would transfect the cells and add the PMA after 2h.
Is a quite simple transfection method and always resulted, as I confirmed by qPCR the transfection.
https://www.vodenmedical.com/images/vodenmedical/lifescience/Final_LabBook_B-G_print.pdf
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