293T suspensions cells were first cultured in a big bubble-stirred bioreactors for large-scale lentivirus vectors (LV) production in the study. Direct domesticated approaches were utilized for 293 suspensions cells. We validated of the ability to LV package and drew a growth curve in shake flask. The results show 293T suspension culture was successful, and also retained the capacity for LV production. After facile optimization of the viral package and cryopreservation conditions, 293T suspension cells were inoculated in a 5.5 L bioreactor. Packaged CAR-CD30 lentivirus yield was close to that from the original shake flask culture. Cultivation for 4 days in the bioreactor enabled production of 1.5±0.3×107 TU/mL coarse LV and the lentiviral infection efficiency was 48.6±2.8% in T Cells. The process has merits of low cost as well as more effective, and expected to be highly competitive compared to other bioreactors.