Dear Reza!

You see in this article, please:

Article Efficient Transient Transfection of Human Multiple Myeloma C...

Figure 3

Electroporation of INA-6 cells stably expressing enhanced green fluorescent protein with an siRNA oligonucleotide against EGFP.

Hey Reza,

We established a method for efficient transient transfection with the JetMessenger transfection reagent and in vitro transcribed and mRNA. This method was used for in vitro transfection of ex vivo (peritoneal macrophages), in vitro (bone marrow-derived macrophages) and MEF cells so far. But you can give it a try with your cell types, if you want. We reached transfection efficiancies of 80 - 90 % with MEFs and BMDM and 65-70% with PMs.

For a detailed protocol (including primer desing, kits of choice and purification of mRNA) please have a look at:

Highly Efficient Transfection of Primary Macrophages with In Vitro Transcribed mRNA. J. Vis. Exp. (Pending Publication), e60143, In-press (2019).

All the best and stay healthy,

Marc

You can always try to package your plasmid DNA into either to lenti or retro viruses since there are plenty of packaging kits are available. Then you can infect those viruses into your primary cells to get higher efficiency of introducing your gene of interest in the primary cells.

That is true. We were forced to use mRNA, because macrophages, as immune cells go into cell death, when foreign DNA is in the cytosol, especially plasmid DNA. But for all other cell types this is the most efficient method.