Transfection of pCI-neo plasmid into 3T3-L1 cells - (Oct/12/2012 )

Hi all

I'm currently transfecting a pCI-neo plasmid with my gene of interest into 3T3-L1 preadipocytes and have a few questions.

the transfection protocol suggests cells should be at 70-90% before transfection and transfection time should be 18-72 hours.

I was planning to do 48 hour transfection time but I'm worried about cells becoming confluent during that time and losing proliferation potential.

I was wondering if I should reduce the level confluency at transfection (maybe 40-50%) to prevent cell reaching confluency. also I was wondering what others have done when transfected into 3T3-L1.


thanks in advance

You can lower the confluency, there is no problem with that. 70% is typically optimal in terms of transfection due to the stuff that cells secrete and other cells need to keep them happy.