Establishing functional lentiviral vector production in a stirred bioreactor for CAR-T cell therapy.
Bioengineered
; 12(1): 2095-2105, 2021 12.
Article
em En
| MEDLINE
| ID: mdl-34047682
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 × 107 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.
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1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Cultura de Vírus
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Imunoterapia Adotiva
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Lentivirus
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Reatores Biológicos
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Vetores Genéticos
Limite:
Humans
Idioma:
En
Revista:
Bioengineered
Ano de publicação:
2021
Tipo de documento:
Article