Cationic mRNA Lipid Nanoparticles for Ex Vivo NanoCAR-T Cell Engineering - PubMed
3 hours ago
- #lipid nanoparticles
- #CAR-T cells
- #mRNA delivery
- Lipid nanoparticles (LNPs) are a non-viral alternative for mRNA-based CAR-T cell engineering.
- Charge-neutral C12-200 LNPs and cationic DOTAP(C12-200) LNPs were compared for ex vivo transfection of primary human T cells.
- C12-200 LNPs achieved efficient transfection with high cell viability in serum-free medium supplemented with apolipoprotein E.
- Transfection efficiency correlated with low-density lipoprotein receptor expression following T cell activation.
- Cationic LNPs demonstrated superior mRNA expression independent of medium composition or T cell activation state but with reduced cytocompatibility.
- Both LNPs efficiently delivered anti-CD20 nanoCAR mRNA into primary T cells, enabling potent cytotoxicity against CD20+ Raji cells in vitro.
- LNP charge, selected proteins in the culture medium, and T cell activation collectively play a critical role in ex vivo T cell engineering.
- Cationic LNPs offer unspecific charge-dependent transfection, inducing high mRNA expression independent of T cell activation status or complex cell culture media.