Phorbol 12-myristate 13-acetate (PMA) is used to differentiate human THP-1 monocytes for 48 hr. into premature macrophages prior to transfection with siRNA.
For transfection the pre-differentiated macrophages are detached enzymatically by Accutase I treatment. The transfection is performed using a Nucleofector 2b device for electroporation of the cells. After transfection, differentiation is continued for another 24 to 48 hr. as required. Finally, mature transfected macrophages are incubated with different kinds of compounds for functional studies.
For more information on the protocol please refer to the research article below. It will be helpful.
Using this protocol for transfection of THP-1 macrophages with siRNA transfection rates of above 90% was achieved without significant reduction of cell vitality.
Hi Malcolm Nobre thanks for your answer. Have you tried transfecting them when they are monocytes? Also, I do not have the Lonza Instrument. I was hoping to get parameters for monocytes on the Neon.