should I use transient or stable transfection - (Aug/20/2007 )

HI,
I am studying the effects of expressing a particular growth factor receptor on the differentiation progess of chondrocytes.
The problem is that the differentiation process takes 5-12 days and if I do a transient transfection I am worried that the effects of my receptor on the differentiation process will be lost after 4-5 days. However, if I make a stable cell line, I think after period of 2-3 weeks expressing the receptor (during the selection period etc) I will not get a chance to see the effect on differentiation of introducing my receptor.

ps. It is the receptor itself I want to study the effects of, it is a mutant form and is constitutively active, and ligand dependent.

Any thought would be much appreciated!

I'd recommend thinking about a stable transfected cell line but with a Tet on/off plasmid. That way you could make the stable lines and then induce expression when you need it. Would save you a lot of work in the long run if you intend to do more than just one experiment.

I would agree with leserattePD that tet on or off system would help you for this sort of experiments. I am a bit cautious using these systems as most old ones are leaky . You might have to get a new one which is not leaky.

I think that the leaky problem is primarily with TetOn. The TetOff system should be fine.