Hek 293f viral production cells

Lentivirus (LV) was produced in HEK 293F viral production cells (Gibco, A35347) using the LV-MAX Transfection Kit (Gibco, A35346) exactly as recommended by the manufacturer using the 30 mL shaker flask culture specifications. Second generation packaging system mix (abm, LV003) was used along with the synthesized SARS-CoV-2 Spike LV transfer vector. Harvest and concentration of the produced LV was performed exactly as recommended by the manufacturer of the LV-MAX Transfection Kit.

HEK 293F viral production cells (Gibco, A35347) were commercially obtained and used for production of lentivirus. Cells were maintained in LV-MAX Production Medium (Gibco, A3583401) in shaker flasks shaking at 135 RPM in a 37°C incubator at 8% CO₂. Cells were subcultured every 2–3 days as per recommendation by the supplier. HEK 293F viral production cells were not authenticated in house. HEK 293F viral production cells were tested and confirmed negative for mycoplasma contamination.
FreeStyle 293F cells (ThermoFisher, R79007) were commercially obtained. Cells were maintained in FreeStyle 293 Expression Medium (ThermoFisher, 12338018) in shaker flasks shaking at 135 RPM in a 37°C incubator at 8% CO₂. Cells were subcultured every 2–3 days as per recommendation by the supplier. FreeStyle 293F cells were not authenticated in house. FreeStyle 293F cells were tested and confirmed negative for mycoplasma contamination.