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Retroviral transduction of mouse t lymphocytes - (Jul/11/2005 )

I´m trying to transduce mouse lymphocytes using a packaging cell line called GP+E86. I´ve tried a lot of things (centrifugation with viral supernatant, coculture of the mouse T-cells with the packaging cell line, and at different time points, and using polybrene all the time). At the moment, the best result that I obtained is 7% (measuring the GFP by flow cytometry), and actually I´m also not so sure about that 7% since they are not very bright and maybe is just a problem of my instrument settings. Has anyone experience in transducing mouse T cells? Should I try something different?
Any suggestions are welcome wink.gif
Thank you smile.gif


i've observed better results with decomplemented serum (just heat it 56° for 30').


I believe retro needs cells to be cycling, are your cells cycling? You may want to stimulate them.