Dear all,
can anyone suggest me a MOUSE cell-line which is highly efficient transfected using normal transfection reagents (e.g. Lipofectamine-3000, XtremeGene HP, FuGENE HD) beside N2a (Neuo-2a). I alredy tested spontaneously immortalized MEFs (3T3 method) which are not really efficient to transfect.
For human (HeLa, HEK, U2-OS, etc.) there are many but not for mouse, it seems to me.
The efficiency should be in the range of N2a, HEK, HeLA etc. (>80-90% transfection efficiency)
Many thanks in advance!
I would try NIH 3T3 cells, ATCC CRL-1658. They have been the best transfecting of a few different 3T3 type lines that I've tried. They transfected well for me using Lipofectamine 2000, but not as high as you state (upwards of 50% by EGFP expression from HCMV promoter/enhancer). The Thermo page for Lipofectamine 3000 however claims that this reagent gives 4-fold higher efficiency (for protein expression) than the 2000 reagent. My guess is that you might get NIH 3T3 cells up to 75 - 80% transfected with Lipofectamine 3000 if the conditions are optimized.
Good luck!
https://www.thermofisher.com/us/en/home/brands/product-brand/lipofectamine/lipofectamine-3000.html
Also, I just remembered that we got transfection efficiencies of 80 - 90% using mouse fibroblast lines that were immortalized with SV40 large T antigen. By transfecting with plasmids containing SV40 ori and HCMV promoter/enhancer expression of the transgene, a colleague was able to optimize FuGene 6 transfection to yield those very high efficiencies. His lab either made the line or they got it as a gift (they made one line and got the other as a gift), and a quick survey of ATCC shows a similar line called SYF, but I don't see any % efficiency data after a quick survey.
Many thanks for you answer! I was also thinking of the NIH 3T3 cells but since I have also spontaneously immortalized MEFs, I thought, there wouldn't be any big differences. > 50% is already also quite okay for my purposes. I will give NIH 3T3 cells a try (have to first ask around in my institute).
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