Influence of γ-secretase inhibition by DAPT on human cellularity, proliferative status, and annexin V+ cells in cell subsets in FTOC seeded with CD34+CD1- thymic progenitor cells after different time points of culture
Cell type, treatment . | Day 4 . | Day 7 . | Day 10 . | Day 20 . | Day 26 . |
|---|---|---|---|---|---|
| Cell no. | |||||
| Control | 16 391 ± 2 911 | 49 107 ± 315 | 61 303 ± 9 024 | 502 068 ± 13 762 | 828 750 |
| 2 μM DAPT | 11 389 ± 713 | 56 108 ± 4 024 | 108 889 ± 45 035 | 285 806 ± 43 782 | 148 353 |
| 5 μM DAPT | 13 016 ± 2 674 | 40 626 ± 5 763 | 123 874 ± 5 633 | 94 166 ± 75 717 | 38 500 |
| S/G2/M cells in total cell population, % | |||||
| Control | 17 ± 5.9 | 10 ± 1.4 | 12 ± 4.2 | 18 ± 2.4 | 6.2 |
| 2 μM DAPT | 22 ± 7.4 | 18 ± 4.2 | 20 ± 2.8 | 4 ± 1.2 | 1.3 |
| 5 μM DAPT | 25 ± 1.3 | 27 ± 0.8 | 15 ± 0.4 | 1 ± 0.1 | 0.5 |
| Annexin V+ cells in total cell population, % | |||||
| Control | 45 ± 12.7 | 42 ± 5.7 | 55 ± 3.5 | 38 ± 6.4 | 32 |
| 2 μM DAPT | 49 ± 14.4 | 37 ± 1.3 | 38 ± 2.0 | 32 ± 8.4 | 36 |
| 5 μM DAPT | 52 ± 16.5 | 49 ± 5.4 | 44 ± 9.1 | 64 ± 9.3 | 65 |
| S/G2/M cells in CD4+ subset, % | |||||
| Control | ND | 8 | 9 ± 2.3 | 23 ± 2.1 | 6.2 |
| 2 μM DAPT | ND | 20 | 18 ± 3.1 | 8 ± 2.6 | 1.3 |
| 5 μM DAPT | ND | 27 | 15 ± 0.6 | 2 ± 0.7 | 0.5 |
| Annexin V+ cells in CD4+ subset, % | |||||
| Control | ND | 40 ± 1.8 | 55 ± 3.6 | 51 ± 10.5 | 44 |
| 2 μM DAPT | ND | 36 ± 4.0 | 38 ± 4.6 | 41 ± 12.6 | 48 |
| 5 μM DAPT | ND | 48 ± 0.1 | 49 ± 9.2 | 66 ± 11.9 | 73 |
| S/G2/M cells in CD4+CD8+ subset, % | |||||
| Control | ND | ND | ND | 12 ± 1.7 | 3.6 |
| 2 μM DAPT | ND | ND | ND | 1 ± 0.3 | 0.5 |
| 5 μM DAPT | ND | ND | ND | 1 ± 0.3 | 0.2 |
| Annexin V+ cells in CD4+CD8+ subset, % | |||||
| Control | ND | ND | ND | 42 ± 12.6 | 41 |
| 2 μM DAPT | ND | ND | ND | 43 ± 11.0 | 51 |
| 5 μM DAPT | ND | ND | ND | 69 ± 15.8 | 74 |
Cell type, treatment . | Day 4 . | Day 7 . | Day 10 . | Day 20 . | Day 26 . |
|---|---|---|---|---|---|
| Cell no. | |||||
| Control | 16 391 ± 2 911 | 49 107 ± 315 | 61 303 ± 9 024 | 502 068 ± 13 762 | 828 750 |
| 2 μM DAPT | 11 389 ± 713 | 56 108 ± 4 024 | 108 889 ± 45 035 | 285 806 ± 43 782 | 148 353 |
| 5 μM DAPT | 13 016 ± 2 674 | 40 626 ± 5 763 | 123 874 ± 5 633 | 94 166 ± 75 717 | 38 500 |
| S/G2/M cells in total cell population, % | |||||
| Control | 17 ± 5.9 | 10 ± 1.4 | 12 ± 4.2 | 18 ± 2.4 | 6.2 |
| 2 μM DAPT | 22 ± 7.4 | 18 ± 4.2 | 20 ± 2.8 | 4 ± 1.2 | 1.3 |
| 5 μM DAPT | 25 ± 1.3 | 27 ± 0.8 | 15 ± 0.4 | 1 ± 0.1 | 0.5 |
| Annexin V+ cells in total cell population, % | |||||
| Control | 45 ± 12.7 | 42 ± 5.7 | 55 ± 3.5 | 38 ± 6.4 | 32 |
| 2 μM DAPT | 49 ± 14.4 | 37 ± 1.3 | 38 ± 2.0 | 32 ± 8.4 | 36 |
| 5 μM DAPT | 52 ± 16.5 | 49 ± 5.4 | 44 ± 9.1 | 64 ± 9.3 | 65 |
| S/G2/M cells in CD4+ subset, % | |||||
| Control | ND | 8 | 9 ± 2.3 | 23 ± 2.1 | 6.2 |
| 2 μM DAPT | ND | 20 | 18 ± 3.1 | 8 ± 2.6 | 1.3 |
| 5 μM DAPT | ND | 27 | 15 ± 0.6 | 2 ± 0.7 | 0.5 |
| Annexin V+ cells in CD4+ subset, % | |||||
| Control | ND | 40 ± 1.8 | 55 ± 3.6 | 51 ± 10.5 | 44 |
| 2 μM DAPT | ND | 36 ± 4.0 | 38 ± 4.6 | 41 ± 12.6 | 48 |
| 5 μM DAPT | ND | 48 ± 0.1 | 49 ± 9.2 | 66 ± 11.9 | 73 |
| S/G2/M cells in CD4+CD8+ subset, % | |||||
| Control | ND | ND | ND | 12 ± 1.7 | 3.6 |
| 2 μM DAPT | ND | ND | ND | 1 ± 0.3 | 0.5 |
| 5 μM DAPT | ND | ND | ND | 1 ± 0.3 | 0.2 |
| Annexin V+ cells in CD4+CD8+ subset, % | |||||
| Control | ND | ND | ND | 42 ± 12.6 | 41 |
| 2 μM DAPT | ND | ND | ND | 43 ± 11.0 | 51 |
| 5 μM DAPT | ND | ND | ND | 69 ± 15.8 | 74 |
Results represent mean ± SEM of 2 independent experiments, except for figures without SEM shown, when only 1 experiment was performed.
ND indicates not done.