HEK293FT or T cells are very easily detached from cell culture vessels, several LV production protocols suggest use poly-d-lysine to coat the culture vessels. Can I avoid coating step when I use Lipofectamine 3000 to do the LV production?
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Yes, you can avoid the poly-d-lysine coating step when using Lipofectamine 3000 as your transfection reagent to produce LV lentivirus.
The protocol for Lipofectamine 3000 LV production requires high cell density, 95-99% confluence at the time of transfection. This prevents HEK293T (or FT) cells from detaching from the culture vessel and causing any interruption due to adding DNA/reagent complex, changing media, harvest LV by replacing fresh media.
It is still important to carefully handle the cells when adding complexes and changing media. It is recommended that any addition should be done towards the side of the culture vessel, and also use pre-warmed media.
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