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Establishing functional lentiviral vector production in a stirred bioreactor for CAR-T cell therapy.
- Source :
-
Bioengineered [Bioengineered] 2021 Dec; Vol. 12 (1), pp. 2095-2105. - Publication Year :
- 2021
-
Abstract
- As gene delivery tools, lentiviral vectors (LV) have broad applications in chimeric antigen receptor therapy (CAR-T). Large-scale production of functional LV is limited by the adherent, serum-dependent nature of HEK293T cells used in the manufacturing. HEK293T adherent cells were adapted to suspension cells in a serum-free medium to establish large-scale processes for functional LV production in a stirred bioreactor without micro-carriers. The results showed that 293 T suspension was successfully cultivated in F media (293 CD05 medium and SMM293-TII with 1:1 volume ratio), and the cells retained the capacity for LV production. After cultivation in a 5.5 L bioreactor for 4 days, the cells produced 1.5 ± 0.3 × 10 <superscript>7</superscript> TU/mL raw LV, and the lentiviral transduction efficiency was 48.6 ± 2.8% in T Cells. The yield of LV equaled to the previous shake flask. The critical process steps were completed to enable a large-scale LV production process. Besides, a cryopreservation solution was developed to reduce protein involvement, avoid cell grafting and reduce process cost. The process is cost-effective and easy to scale up production, which is expected to be highly competitive.
Details
- Language :
- English
- ISSN :
- 2165-5987
- Volume :
- 12
- Issue :
- 1
- Database :
- MEDLINE
- Journal :
- Bioengineered
- Publication Type :
- Academic Journal
- Accession number :
- 34047682
- Full Text :
- https://doi.org/10.1080/21655979.2021.1931644