What do people commonly get as a transfection efficiency using plasmids for commonly used AML related suspension cell lines? Is 50% good? Does anyone have any special tips and tricks?
we usually don't transfect plasmids directly to these AML cell lines, we use lentivirus to infect U937 cells, which can get a more than 90% efficiency.
From my own experience with Viromers (polymer-based reagents) and other chemical transfectants, there is a strong limitation for transfecting plasmids in these cells. It's commonly assumed that enzymatic DNAse reactions are activated in response to foreign DNA uptake. A great alternative is to transfect the corresponding transcript instead (if possible modified mRNA obtained by IVT or from an oligo supplier). But to answer your question, 50% is quite good, I would rather hope for max 20-30%.
For sure, there is no limitation for these cells to uptake the transfection complex (as shown with great efficiency obtained for siRNA or mRNA transfection), but with DNA, further processing after intracellular delivery is the issue.
Bests, Sandra
https://www.researchgate.net/institution/Lipocalyx_Gmbh/posts
- Badges
- Science method