Lentivirus transduction of spleen cells - (May/20/2009 )
I am transfecting 293T cells with packaging lentiviral plasmids to generate a lentiviral particles carrying GFP and another gene. When I infect 293T cells with the viral particles to calculate the viral titer I get a very high efficient GFP signal on FACS. But the problem is, when I infect mice spleen cells nothing really happens !. Anybody ever used lentivirus for transduction of cells ? Do you have any suggestions to imprvoe the infection of the cells?
Lentivirus are a pretty strong way to infect cells. That said, I don't think you can compare 293T cells (whitch you could infect by simply looking at them since they are so easy to infect/transfect) and live spleen cells for virus efficiency. You may try to add more virus and check afterwards. It's also important to add 8µg/ml of polybrene to your virus : it'll greatly enhance the virus's capacity to bind and infected cells.
You are also crossing species -293 are human, mouse spleen is, well..., mouse. This may have some affect on how well the lentivirus works (I've never used them so I don't know, but typically viruses are reasonably species specific).