Main Characteristics of Injected CB CD34+Cells and Their Ability to Engraft the BM of NOD/SCID Recipients
| Injected/Mouse . | No. of Positive Mice . | Engraftment (%) . | Human CFC (×103)* . | ||
|---|---|---|---|---|---|
| CD34+ Cells . | CFC . | LTC-IC . | |||
| 2 × 105 | 49,330 | 17,800 | 3/3† | 12.5, 21, 7‡ | 51.2, 98.4, 27.6 |
| 1 × 105 | 24,665 | 8,900 | 4/4 | 1.3, 3, 11, 25 | 17.7, 14.2, 41.5, 77.6 |
| 5 × 104 | 12,332 | 4,450 | 4/5 | 1.4, 16, 3, 8, 0 | ND, 74.9, 10.2, 36.4, 0 |
| 3.5 × 104 | 8,631 | 3,115 | 4/6 | 1.2, 2.3, 0, 0, 1.6, 3 | 4.9, 9.8, 0, 0, 8.08, 9.5 |
| 2 × 104 | 4,933 | 1,780 | 3/7 | 0.18, 0.5, 0.3, 0, 0, 0, 0 | 0.76, 2.1, 1.9, 0, 0, 0, 0 |
| 1 × 104 | 2,466 | 890 | 0/5 | 0, 0, 0, 0, 0 | 0, 0, 0, 0, 0 |
| Injected/Mouse . | No. of Positive Mice . | Engraftment (%) . | Human CFC (×103)* . | ||
|---|---|---|---|---|---|
| CD34+ Cells . | CFC . | LTC-IC . | |||
| 2 × 105 | 49,330 | 17,800 | 3/3† | 12.5, 21, 7‡ | 51.2, 98.4, 27.6 |
| 1 × 105 | 24,665 | 8,900 | 4/4 | 1.3, 3, 11, 25 | 17.7, 14.2, 41.5, 77.6 |
| 5 × 104 | 12,332 | 4,450 | 4/5 | 1.4, 16, 3, 8, 0 | ND, 74.9, 10.2, 36.4, 0 |
| 3.5 × 104 | 8,631 | 3,115 | 4/6 | 1.2, 2.3, 0, 0, 1.6, 3 | 4.9, 9.8, 0, 0, 8.08, 9.5 |
| 2 × 104 | 4,933 | 1,780 | 3/7 | 0.18, 0.5, 0.3, 0, 0, 0, 0 | 0.76, 2.1, 1.9, 0, 0, 0, 0 |
| 1 × 104 | 2,466 | 890 | 0/5 | 0, 0, 0, 0, 0 | 0, 0, 0, 0, 0 |
CD34+ cells deriving from 6 different CB samples were separated as described, pooled, and then stored frozen. At the time of injection, the samples were thawed, washed, counted, resuspended in 300 μL of IMDM containing 10% FCS and then injected into 3 to 7 animals as described. BM of NOD/SCID recipients was recovered from both the femurs and the tibias at week 8 posttransplant.
Abbreviation: ND, not done.
A total of 1 × 105 to 5 × 105 BM cells of the transplanted mice were seeded in triplicate dishes for plasma clot assays. Recombinant human IL-3, GM-CSF, SCF, and EPO were added at start of cultures. Dishes were scored after 14 days of incubation, after staining with MoAb as described in Materials and Methods. Only fluorescent colonies were considered of human origin. Values are calculated taking into account that the BM of both femurs and tibias represent 25% of the total BM.
Positive mouse = >0.1% of human CD45+, CD71+, and α-GpA A+ cells in the whole BM on FACSscan analysis.
Percent of human CD45+, CD71+, and GpA+ cells in the whole BM on FACSscan analysis.