Optimized High-Titer Lentivirus Production and Efficient CAR-T Cell Generation - PubMed
5 hours ago
- #lentivirus production
- #CAR-T cells
- #cancer immunotherapy
- CAR-T cell therapy has revolutionized cancer immunotherapy, especially for B-cell malignancies, with high response rates in acute lymphoblastic leukemia.
- Emerging applications of CAR-T cells include treating autoimmune diseases like lupus, showcasing their broad therapeutic potential.
- Manufacturing CAR-T cells, particularly producing high-titer lentiviral vectors (LV), remains a significant challenge, limiting accessibility and ease of production.
- Optimized protocols for LV production include adjusting plasmid ratios, enhancing transfection efficiency in HEK293T cells, and fine-tuning PBMC activation and transduction conditions.
- The study presents scalable protocols for third-generation LV production to facilitate reproducible and cost-effective CAR-T cell generation for preclinical and translational research.
- Key protocols include plasmid ratio screening, viral concentration via ultracentrifugation, FACS-based titration, PBMC isolation and activation, and CAR-T cell generation with lentivirus.