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Establishing 293T suspension cultrue for lentiviral vector production in a stirred bioreactor
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  • Qulai Tang,
  • Lixing Gu,
  • Han Wu,
  • Fan Li,
  • Haichuan Zhu,
  • Chaojiang Gu,
  • Shangkun Zhang,
  • Jinjue Liang,
  • Wei Qi,
  • Yong Zhou,
  • Tongcun Zhang
Qulai Tang
Tianjin University of Science and Technology
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Lixing Gu
Wuhan University of Science and Technology
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Han Wu
Wuhan University of Science and Technology
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Fan Li
Wuhan University of Science and Technology
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Haichuan Zhu
Wuhan University of Science and Technology
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Chaojiang Gu
Wuhan University of Science and Technology
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Shangkun Zhang
Wuhan University of Science and Technology
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Jinjue Liang
Wuhan University of Science and Technology
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Wei Qi
Tianjin University of Science and Technology
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Yong Zhou
Wuhan University of Science and Technology
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Tongcun Zhang
Tianjin University of Science and Technology
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Abstract

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.

Peer review status:POSTED

10 Nov 2020Submitted to Biotechnology and Bioengineering
10 Nov 2020Assigned to Editor
10 Nov 2020Submission Checks Completed