RDTR/SCFHA-LV long-term in vivo transduction of hHSCs in humanized mice
| Mice . | Reconstitution, % hCD45+ cells* . | BM, % GFP+ cells/total Lin+ cells† . | Thymus, % GFP+ cells† . | Spleen, % GFP+ cells† . | PB, % GFP+ cells† . | CFCs, % GFP+ cells‡ . | |||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| BM . | Thy . | Spl . | PB . | h45 . | h34 . | h13 . | h19 . | h45 . | h45 . | h45 . | |||
| RDTR/SCFHA 1 | 18 | 98 | 24 | 9 | IF1 + 2 | 4 | 3.9 | 4 | 1.7 | 0 | 0.1 | 0 | 4.6 |
| RDTR/SCFHA 2 | 25 | 97 | 21 | 5 | IF1 + 2 | 2.4 | 3.6 | 3 | 0.2 | 0 | 0 | 0 | ND |
| RDTR/SCFHA 3 | 10 | 98.3 | 20 | 1 | IF1 + 2 | 3.4 | 5.7 | 7.1 | 1.1 | 0 | 0.3 | 0 | 4.3 |
| RDTR/SCFHA 4 | 5 | 97 | 10 | 2 | IF1 + 2 | 5.5 | 4 | 5.4 | 0.8 | 0 | 0.4 | 0 | 5.5 |
| RDTR/SCFHA 5 | 16 | 91 | 49 | 6 | IF1 + 2 | 3.7 | 5.5 | 5.3 | 1.6 | 0 | 0.4 | 0 | 5.3 |
| RDTR/SCFHA 6 | 7 | 98 | 38 | ND | IF1 + 2 | 3.4 | 4.9 | 4.1 | 1.4 | 0 | 0.1 | 3.8 | |
| RDTR/SCFHA 7 | 5 | 99 | 42 | 2 | IF1 + 2 | 3 | 6.4 | 8.9 | 0.5 | 0 | 0.03 | 0 | 5.8 |
| Mice . | Reconstitution, % hCD45+ cells* . | BM, % GFP+ cells/total Lin+ cells† . | Thymus, % GFP+ cells† . | Spleen, % GFP+ cells† . | PB, % GFP+ cells† . | CFCs, % GFP+ cells‡ . | |||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| BM . | Thy . | Spl . | PB . | h45 . | h34 . | h13 . | h19 . | h45 . | h45 . | h45 . | |||
| RDTR/SCFHA 1 | 18 | 98 | 24 | 9 | IF1 + 2 | 4 | 3.9 | 4 | 1.7 | 0 | 0.1 | 0 | 4.6 |
| RDTR/SCFHA 2 | 25 | 97 | 21 | 5 | IF1 + 2 | 2.4 | 3.6 | 3 | 0.2 | 0 | 0 | 0 | ND |
| RDTR/SCFHA 3 | 10 | 98.3 | 20 | 1 | IF1 + 2 | 3.4 | 5.7 | 7.1 | 1.1 | 0 | 0.3 | 0 | 4.3 |
| RDTR/SCFHA 4 | 5 | 97 | 10 | 2 | IF1 + 2 | 5.5 | 4 | 5.4 | 0.8 | 0 | 0.4 | 0 | 5.5 |
| RDTR/SCFHA 5 | 16 | 91 | 49 | 6 | IF1 + 2 | 3.7 | 5.5 | 5.3 | 1.6 | 0 | 0.4 | 0 | 5.3 |
| RDTR/SCFHA 6 | 7 | 98 | 38 | ND | IF1 + 2 | 3.4 | 4.9 | 4.1 | 1.4 | 0 | 0.1 | 3.8 | |
| RDTR/SCFHA 7 | 5 | 99 | 42 | 2 | IF1 + 2 | 3 | 6.4 | 8.9 | 0.5 | 0 | 0.03 | 0 | 5.8 |
Two- to 4-day-old newborn BALB/c Rag2−/−γc−/− mice were subjected to a sublethal irradiation of 2 × 1.5 Gy. 2 × 105 hCB-CD34+ cells were injected intrahepatically into the newborns. After 9 weeks of reconstitution, the humanized mice were injected into both femurs with 10 μL of the RDTR/SCFHA-LVs (5E7–1E8 IU/mL). The animals were killed 6-8 weeks after intrafemoral injection of the vectors and the BM, spleens, thymi, and PB were harvested and assessed for levels of human engraftment and GFP-expressing human cells. Independent experiments were performed with different CD34+ CB samples and different preparations of each vector.
ND indicates not done.
Percentage of human cells (hCD45+ cells) per total human and mouse mononucleated cells in each hematopoietic tissue.
Percentages of GFP+ cells per total Lin+ cells (ie, the percentage of GFP+CD13+ cells/total CD13+ cells).
Six to 8 weeks after RDTR/SCFHA-LV intrafemoral injection into these humanized mice, hCD34+ cells were isolated from part of the humanized BM by magnetic cell isolation (Miltenyi Biotec) and then seeded in methyl cellulose to induce myeloid differentiation and to score the percentage of GFP+ CFCs.