Transduction efficacy of mobilized peripheral blood CD34+ cells from healthy donors exposed to a short transduction with a GMP-produced PGK-FANCA.Wpre* lentiviral vector
| . | . | Total cells (×106) . | Viability . | Total colonies (×105) . | Colonies/1000 CD34+ cells . | VCN/cell (14-d liquid culture) . |
|---|---|---|---|---|---|---|
| Validation I | Input | 3.00 | 90.96 | 4.73 | 158 | — |
| Output | 1.10 (36.7%) | 83.60 (91.91) | 1.30 (27.4%) | 118 (74.7%) | 0.39 | |
| Validation II | Input | 3.00 | 88.76 | 5.63 | 188 | — |
| Output | 1.56 (51.0%) | 94.03 (105.94) | 2.30 (47.4%) | 150 (79.9%) | 0.33 | |
| Validation III | Input | 3.00 | 86.4 | 7.63 | 254 | — |
| Output | 1.39 (46.3%) | 96.3 (111.46) | 3.45 (44.9%) | 247 (96.9%) | 0.67 | |
| Mean ± SE | Input | 3.00 ± 0 | 88.71 ± 1.32 | 6.00 ± 0.86 | 200 ± 28.35 | — |
| Output | 1.34 ± 0.13 (44.7% ± 1.92%) | 91.31 ± 3.91 (103.10 ± 5.82) | 2.34 ± 0.62 (37.7% ± 6.29%) | 171 ± 38.78 (83.8% ± 6.94%) | 0.40 ± 0.10 |
| . | . | Total cells (×106) . | Viability . | Total colonies (×105) . | Colonies/1000 CD34+ cells . | VCN/cell (14-d liquid culture) . |
|---|---|---|---|---|---|---|
| Validation I | Input | 3.00 | 90.96 | 4.73 | 158 | — |
| Output | 1.10 (36.7%) | 83.60 (91.91) | 1.30 (27.4%) | 118 (74.7%) | 0.39 | |
| Validation II | Input | 3.00 | 88.76 | 5.63 | 188 | — |
| Output | 1.56 (51.0%) | 94.03 (105.94) | 2.30 (47.4%) | 150 (79.9%) | 0.33 | |
| Validation III | Input | 3.00 | 86.4 | 7.63 | 254 | — |
| Output | 1.39 (46.3%) | 96.3 (111.46) | 3.45 (44.9%) | 247 (96.9%) | 0.67 | |
| Mean ± SE | Input | 3.00 ± 0 | 88.71 ± 1.32 | 6.00 ± 0.86 | 200 ± 28.35 | — |
| Output | 1.34 ± 0.13 (44.7% ± 1.92%) | 91.31 ± 3.91 (103.10 ± 5.82) | 2.34 ± 0.62 (37.7% ± 6.29%) | 171 ± 38.78 (83.8% ± 6.94%) | 0.40 ± 0.10 |
A total of 3 × 106 CD34+ cells were prestimulated in 3 mL of supplemented medium for 8 to 10 hours, followed by another 12 to 14 hours of transduction with the therapeutic lentiviral vector. Analyses of VCN/cell was conducted in cells maintained in liquid culture for 14 days, according to a validated GMP protocol. Percentages (in parentheses) determined after transduction (output) with respect to values determined before transduction (input) are shown. In these experiments, CD34+ cells were mobilized with G-CSF.