primary valve interstitial cells transfection - (Jan/15/2013 )
I want to do transient transfection of valve interstitial cells isolated from pig aortic valves. I can't use viral vector, and at first I really need to try transfection by some reagent (after that I'll have opportunity to do this by electroporation). Can you suggest me any thing? I've try to use PEI but I can't see any effect (EGFP vector). Can you suggest anything?
There are a lot of different reagents, but the two most common are Fugene and Lipofectamine. Fugene and its derivatives tend to work well with low toxicity. Lipofectamine and its derivatives are OK but tend to have a lot of toxicity. PEI is a base for most of them but many of the variations will be better at transfecting primary cell lines, which tend to be difficult to transfect.
Have you tested that your GFP vector works with other cell lines?