Efficient transduction of in vivo repopulating HSCs with cyclin D2–containing vector
. | 1° recipients . | . | 2° recipients . | . | ||
|---|---|---|---|---|---|---|
. | % Donor (CD45.2) reconstitution (SD) . | % GFP+ donor cells (SD) . | % Donor (CD45.2) reconstitution (SD) . | % GFP+ donor cells (SD) . | ||
| Experiment 1 | ||||||
| Control vector | 72.7 (5.2) | 48.5 (7.4) | 60.7 (21.0) | 42.5 (42.8) | ||
| Cyclin D2 vector | 76.2 (3.3) | 33.7 (10.6) | 81.9 (10.6) | 36.8 (33.3) | ||
| Experiment 2 | ||||||
| Control vector | 58.4 (12.1) | 99.4 (0.9) | 34.6 (43.3) | 99.1 (1.2) | ||
| Cyclin D2 vector | 43.7 (15.4) | 97.3 (2.9) | 23.0 (18.9) | 98.1 (3.3) | ||
. | 1° recipients . | . | 2° recipients . | . | ||
|---|---|---|---|---|---|---|
. | % Donor (CD45.2) reconstitution (SD) . | % GFP+ donor cells (SD) . | % Donor (CD45.2) reconstitution (SD) . | % GFP+ donor cells (SD) . | ||
| Experiment 1 | ||||||
| Control vector | 72.7 (5.2) | 48.5 (7.4) | 60.7 (21.0) | 42.5 (42.8) | ||
| Cyclin D2 vector | 76.2 (3.3) | 33.7 (10.6) | 81.9 (10.6) | 36.8 (33.3) | ||
| Experiment 2 | ||||||
| Control vector | 58.4 (12.1) | 99.4 (0.9) | 34.6 (43.3) | 99.1 (1.2) | ||
| Cyclin D2 vector | 43.7 (15.4) | 97.3 (2.9) | 23.0 (18.9) | 98.1 (3.3) | ||
Lethally irradiated mice (1° recipients, CD45.1) received transplants with 5000 (experiment 1) or 1250 (experiment 2) LSK (CD45.2) cells transduced with control- or cyclin D2–containing vectors ('Materials and methods'), and 200 000 (experiment 1) or 100 000 (experiment 2) whole BM (CD45.1) cells. Four months later, half a femur equivalent BM cells from 1° recipients were serially transplanted into 2° (CD45.1) recipients. Reconstitution was analyzed in PB 4 months (1° recipients) and 3 months (2° recipients) after transplantation. As shown for 1° recipients in Table 1, control- and cyclin D2–transduced (GFP+) progenitors showed comparable contributions to B (B220), T (CD3), and myeloid (CD11b/Gr1) lineages also in 2° recipients. Data are mean values (SD) from 4 to 7 mice in each group and experiment.