NIH3T3 Transfection - (May/12/2011 )
I need to perform transfection and viral infetion on NIH3T3 for two seperate experiments. I have already done the infection, but it seems the infection efficiency is quite low, compared to other cell lines. NIH3T3 is commonly used for in vitro studies, so it gives me the perception it's easy to transfect and infect. Because of the low infection efficiency, I am a bit worried the same thing will happen on transient transfection using lipofectamine 2000 on NIH3T3. Would you mind sharing your experience and protocol on transfection or infection of this cell line? And are there any tricks? Thanks a lot.
3T3 are indeed rather difficult to transfect or infect. I found that I can increase transfection by 10fold using Calciumphosphate precipitation compared to lipofection. Infection with Lentivirus works rather well (I heard somewhere they would be the "Gold Standard" for this, but in the meantime I found much better cells...). IMHO 3T3 are rather "normal" cells with lots of intact signaling pathways (for example contact inhibition), so they can serve as a good control.