As the clinical application of cell therapeutics advances, the efficient ex vivo expansion of chimeric antigen receptor (CAR) T cells strictly demands optimized, animal-origin-free (AOF) culture systems to ensure uncompromising safety and therapeutic efficacy. In this study, we evaluated a novel AOF media (CellCor™) for CD19 CAR-T cell production. The performance of the AOF media was rigorously confirmed through independent third-party validation conducted by Cytiva (USA), demonstrating objective results. The performance of CellCor was comparatively analyzed against a widely used commercial media, T-VIVO® (Lonza).
Primary CD3+ T cells from three healthy donors were activated (T Cell TransAct™), cultured in either CellCor™ or T-VIVO® with IL-2, and transduced with a CD19-directed lentiviral vector (MOI 2.5). Following a 10-day expansion, cells were evaluated for proliferation, viability, and transduction efficiency. Flow cytometry assessed immunophenotypes (CD3, CD4, CD8, CD25, CD45RO, CD62L, and CD279(PD-1)). In vitro functionality was determined via cytotoxicity assays against CD19+ Nalm-6 cells and ELISA-based cytokine profiling (TNF-α, IFN-γ, Granzyme B, and IL-2).
Both media supported robust expansion with high viability (95–98%). Notably, CellCor™ achieved superior CAR transduction efficiency across all donors (average 56.3% vs. 30.7% for T-VIVO®) with elevated mean fluorescence intensity, indicating stronger CAR surface density. Phenotypically, CellCor™ enriched the CD4+ subset and central memory T cells, whereas T-VIVO® shifted slightly toward CD8+ effector memory phenotypes. Both media exhibited equivalent, potent in vitro cytotoxicity (~80% specific lysis). Furthermore, CellCor™ induced a robust inflammatory cytokine profile (elevated IFN-γ, TNF-α, and IL-2), suggesting advantages for in vivo persistence.
These findings objectively demonstrate that CellCor™ maximizes CAR+ yield and expression while preserving essential cytotoxicity and a favorable central memory phenotype. It serves as a thoroughly validated, highly effective AOF alternative for clinical CAR-T manufacturing.