Figure 2.

Optimization of the lentiviral transduction method for CB CD8⁺ T cells

(A) Transduction efficiency, presented as percentage of GFP⁺ cells, comparing two methods for T cell transduction: spinoculation (black) and overnight incubation (gray) (B) Transduction efficiency, presented as percentage of GFP⁺ cells, comparing the use of different culture media during overnight transduction: X-Vivo15 (gray) and StemMACS media (stripes). (C) Representative flow cytometry plots of methods and media tested using both transduction with PGK.EGFP and MND.EGFP. The density plots are showing the measured CD3 and CD8 expression (on the left) and GFP expression in CD3⁺ CD8⁺ T cells (on the right) in the left (PGK.EGFP) and right (MND.EGFP) panels. (D) Protein expression, presented as MFI of EGFP signal, is statistically increased using the MND promoter. (E) qPCR analysis of the VCN-positive correlates with the MFI results. The data correspond to ≥3 independent experiments and are shown as average ± standard deviation; ∗p ≤ 0.05; ∗∗p ≤ 0.01; ∗∗∗p ≤ 0.001; ∗∗∗∗p ≤ 0.0001.