I am working with rat BM-MSCs, and i am trying to transfect these cell with integrative pIRES-EGFP plasmid by applying lipofectamine 2000 (invitrogen). however I found these cells very hard to transfect (almost 10% efficiency). in addition they seem to lose their proliferation capacity after transfection. does anybody have experience in this field? any suggestion or protocol would be appreciated.
Thankfully. F Shojaei.
This might not be the answer you are looking for but it might save you some time. We gave up on chemical-based transfection of MSCs a few years ago for the same reasons. Nucleofection is an excellent alternative if you can get access to an instrument.
If you look at some papers with H. Uludag as author, you may be able to see transfection rates with Lipo 2000 (
While our Viromer reagents transfect good siRNA, see some examples here
https://viromer-transfection.com/wp-content/uploads/2015/09/mesenchymal-stem-cell-transfection-viromer.pdf
it's much more harder for plasmid transfection.
We have only one reference and it was less than 15% of efficiency, then I confirm they are very difficult to transfect with plasmid.
ref Article Hypoxia Differentially Modulates the Genomic Stability of Cl...
Maybe an alternative would be to use mRNA...
https://viromer-transfection.com/mrna-transfection
Best, Sandra
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