In vitro expansion and in vivo engraftment of lentivirus-transduced cord blood cells
| Cell dose . | T0 . | 1 Week . | 4 Weeks . | |||||||
|---|---|---|---|---|---|---|---|---|---|---|
| Engraftment* . | In vitro . | Engraftment* . | In vitro . | Engraftment* . | ||||||
| CD45+,% . | CD45+/GFP+, % . | Cell no. × 106 (% GFP+cells)† . | CD34+ × 106‡ (% GFP+ cells) . | CD45+, % . | CD45+/GFP+, % . | Cell no. × 106 (% GFP+ cells)† . | CD34+ × 106 (% GFP+ cells)‡ . | CD45+, % . | CD45+/GFP+, % . | |
| 50 000 | ||||||||||
| 3.4 | 9.7 | 1.9 (38) | 0.8 (16) | 6.9 | 8.4 | 9.6 (19.2) | 2.6 (52) | 16.7 | 7.9 | |
| 3.5 | 10.6 | 2.3 (46) | 0.4 (8) | 8.1 | 9.2 | 4.9 (98) | 1.2 (24) | 12 | 8.4 | |
| 4.5 | 12.3 | 1.67 (33.4) | 0.45 (9) | 7.4 | 12.4 | 7.1 (14.2) | 1.9 (38) | 17 | 16.9 | |
| 1.5 | 8.6 | 21.1 (42.2) | ND | ND | ND | 6.8 (13.6) | 0.95 (19) | 11 | 21.3 | |
| 100 000 | ||||||||||
| 8 | 14.7 | 2.1 (21) | 1.4 (14) | 18.4 | 16.4 | 11.6 (11.6) | 2.6 (26) | 26.5 | 12.9 | |
| 12.6 | 9.9 | 7.5 (75) | 1.9 (19) | 26.5 | 10.6 | 36.2 (36.2) | 5.4 (54) | 39 | 8.7 | |
| 13.4 | 12.4 | 4 (40) | 1.3 (13) | 16.0 | 7.5 | 29.2 (29.2) | 4.4 (44) | 21.6 | 10.3 | |
| 9.8 | 11.6 | 5.7 (57) | 2.0 (20) | ND | ND | 18 (18) | 3.3 (33) | 18 | 15.6 | |
| 14.2 | 15.0 | 3.8 (38) | 2.6 (26) | 21 | 14.6 | 30.4 (30.4) | 5.2 (52) | 30.7 | 10.9 | |
| 200 000 | ||||||||||
| 19.5 | 16.8 | 8.9 (44.5) | 2.8 (14) | 27 | 10.6 | 38.4 (19.2) | 10.3 (51.5) | 45.6 | 15.2 | |
| 26.7 | 11.3 | 7.5 (37.5) | 3.1 (15.5) | 29.4 | 14.9 | 44.5 (22.2) | 7.6 (38) | 56.8 | 7.9 | |
| 23.3 | 13.2 | 8.3 (41.5) | 2.9 (14.5) | ND | ND | 63.2 (31.6) | 7.4 (37) | 71.4 | 16.8 | |
| Cell dose . | T0 . | 1 Week . | 4 Weeks . | |||||||
|---|---|---|---|---|---|---|---|---|---|---|
| Engraftment* . | In vitro . | Engraftment* . | In vitro . | Engraftment* . | ||||||
| CD45+,% . | CD45+/GFP+, % . | Cell no. × 106 (% GFP+cells)† . | CD34+ × 106‡ (% GFP+ cells) . | CD45+, % . | CD45+/GFP+, % . | Cell no. × 106 (% GFP+ cells)† . | CD34+ × 106 (% GFP+ cells)‡ . | CD45+, % . | CD45+/GFP+, % . | |
| 50 000 | ||||||||||
| 3.4 | 9.7 | 1.9 (38) | 0.8 (16) | 6.9 | 8.4 | 9.6 (19.2) | 2.6 (52) | 16.7 | 7.9 | |
| 3.5 | 10.6 | 2.3 (46) | 0.4 (8) | 8.1 | 9.2 | 4.9 (98) | 1.2 (24) | 12 | 8.4 | |
| 4.5 | 12.3 | 1.67 (33.4) | 0.45 (9) | 7.4 | 12.4 | 7.1 (14.2) | 1.9 (38) | 17 | 16.9 | |
| 1.5 | 8.6 | 21.1 (42.2) | ND | ND | ND | 6.8 (13.6) | 0.95 (19) | 11 | 21.3 | |
| 100 000 | ||||||||||
| 8 | 14.7 | 2.1 (21) | 1.4 (14) | 18.4 | 16.4 | 11.6 (11.6) | 2.6 (26) | 26.5 | 12.9 | |
| 12.6 | 9.9 | 7.5 (75) | 1.9 (19) | 26.5 | 10.6 | 36.2 (36.2) | 5.4 (54) | 39 | 8.7 | |
| 13.4 | 12.4 | 4 (40) | 1.3 (13) | 16.0 | 7.5 | 29.2 (29.2) | 4.4 (44) | 21.6 | 10.3 | |
| 9.8 | 11.6 | 5.7 (57) | 2.0 (20) | ND | ND | 18 (18) | 3.3 (33) | 18 | 15.6 | |
| 14.2 | 15.0 | 3.8 (38) | 2.6 (26) | 21 | 14.6 | 30.4 (30.4) | 5.2 (52) | 30.7 | 10.9 | |
| 200 000 | ||||||||||
| 19.5 | 16.8 | 8.9 (44.5) | 2.8 (14) | 27 | 10.6 | 38.4 (19.2) | 10.3 (51.5) | 45.6 | 15.2 | |
| 26.7 | 11.3 | 7.5 (37.5) | 3.1 (15.5) | 29.4 | 14.9 | 44.5 (22.2) | 7.6 (38) | 56.8 | 7.9 | |
| 23.3 | 13.2 | 8.3 (41.5) | 2.9 (14.5) | ND | ND | 63.2 (31.6) | 7.4 (37) | 71.4 | 16.8 | |
CB CD34+ cells at the indicated doses were stimulated for 48 hours with FL, TPO, SCF, and IL6; incubated overnight with vector particles (TU 50 × 106/mL, or MOI 50); and injected into sublethally irradiated NOD/SCID mice soon after transduction or after further in vitro culture for 1 or 4 weeks. Results from 3 to 5 different sets of experiments per initial cell dose are shown.
The percentage of human CD45+ cells in total BM cells and the percentage of GFP+ cells within the CD45 population are reported.
Number of cells generated during expansion; in parentheses, the percentage of GFP+ cells within the total cells.
The absolute number of CD34+ cells in the expansion cultures; in parentheses, the percentages of GFP+ cells within the CD34+ population.