I am trying to transduce mouse T cells with CAR constructs using lentivirus. The protocol I am using for now seems inconvienent that transduction is not working at all.

So briefly, I isolate T cells using Miltenyi magentic beads, then I activate them with CD3/CD28 with IL-2 100u/ml for 3 days in complete medium. On day 3, I add the produced viral particles to them in a retronectine coated 12-well plate, then check the transduction effeciency 3 days later.

I use Ecotropic envelop MEE for the viral particles. could anyone help me with a recommendation to improve the T cell transduction?

thanks in advance

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