I would like to differentiate THP-1 cell to macrophage using PMA and transfect pAcGFP vector with Lipofectamine ltx transfection reagent (as it is available).
Is there any standard protocol of transfection of macrophage like THP-1 with lipofectamine? I have found several papers but could not understand completely.
Please share your experience or important reference papers if any. Valuable advice will be appreciated. Thanks in advance.
Hello!
As already discussed on RG, plasmid transfection in most of MPS cells is just limited by their capacity to eliminate foreign DNA once into the cytosol, as part of their role in innate immunity.
With Lipofectamine, I have no reference, but with our polymer-based reagent Viromer RED, our customers achieved never more than 25% of transfected cells with THP-1 monocytes. And it's after tedious optimization, and toxicity is observed.
On the other hand, by transfecting mRNA, 40-50% is reachable, without any sign of toxicity.
If you really want to transfect THP-1 cells, you should really think about switching to mRNA.
Please, have a look to the links below. The first one will give you also some tips for PMA-differentiation used before transfection.
Best, Sandra
https://viromer-transfection.com/wp-content/uploads/2015/09/THP-1-transfection-viromer.pdf
https://www.researchgate.net/institution/Lipocalyx_Gmbh/posts
Hi,
PMA differentiation works very well. I have used it and it worked great (100ng/ml).
I've tried to transfect THP-1 cells with Lipofectamine and it does not work, at least not in my hands.
I transfected my THP-1 cells with the Neon system (electroporation). Here is the PubMed ID PMID:27959965 for my paper where I have described the details.I did it with small interfering RNA but I tested also with plasmids and it worked. You just have to optimize the protocol for plamids.
Hope it helps you :)
Best of luck!
Ghazaleh
- Badges
- Science method