Comparison of the Human CD45+ Cell and Clonogenic Progenitor Content in 1 Million Bone Marrow Cells Recovered From Mice That Received Human CD34+ Progenitor Cells Transduced in Suspension With or Without FL Versus on Stromal Support
| bnx No. . | Human CD45+ Cells per 1 × 106bnx BM Cells . | Human CFU per 1 × 106bnx BM Cells . |
|---|---|---|
| Transplanted with human CD34+ cells transduced in suspension culture with IL-3, IL-6, and SCF | ||
| 43A1 | 3,700 | 03-150 |
| 43A2 | 900 | 0 |
| 44A1 | 03-151 | 0 |
| 44A2 | 1,400 | 0 |
| 44A3 | 0 | 0 |
| 46B1 | 1,100 | 0 |
| 46B2 | 900 | 0 |
| 48A1 | 3,100 | 0 |
| 48A2 | 0 | 0 |
| 48A3 | 0 | 0 |
| Average | 1,110 ± 1,324 | 0 ± 0 |
| Transplanted with human CD34+ cells transduced in suspension culture with IL-3, IL-6, SCF, and FL | ||
| 43B1 | 44,900 | 63 |
| 43B2 | 18,100 | 0 |
| 44B1 | 0 | 10 |
| 44B2 | 44,000 | 103 |
| 44B3 | 40,100 | 153 |
| 46C1 | 53,200 | 177 |
| 46C2 | 31,800 | 20 |
| 46C3 | 73,400 | 107 |
| 48B1 | 29,400 | 103 |
| 48B2 | 75,900 | 57 |
| 48B3 | 0 | 0 |
| Average | 37,345 ± 25,299 | 72 ± 62 |
| Transplanted with human CD34+ cells transduced on stromal support with IL-3, IL-6, and SCF | ||
| 43C1 | 73,200 | 90 |
| 43C2 | 110,800 | 223 |
| 43C3 | 34,200 | 7 |
| 44C1 | 78,900 | 240 |
| 44C2 | 28,200 | 213 |
| 46A1 | 99,100 | 326 |
| 46A3 | 52,500 | 170 |
| 48C1 | 40,900 | 203 |
| 48C2 | 1,500 | 0 |
| 48C3 | 32,000 | 233 |
| Average | 55,130 ± 34,493 | 171 ± 106 |
| bnx No. . | Human CD45+ Cells per 1 × 106bnx BM Cells . | Human CFU per 1 × 106bnx BM Cells . |
|---|---|---|
| Transplanted with human CD34+ cells transduced in suspension culture with IL-3, IL-6, and SCF | ||
| 43A1 | 3,700 | 03-150 |
| 43A2 | 900 | 0 |
| 44A1 | 03-151 | 0 |
| 44A2 | 1,400 | 0 |
| 44A3 | 0 | 0 |
| 46B1 | 1,100 | 0 |
| 46B2 | 900 | 0 |
| 48A1 | 3,100 | 0 |
| 48A2 | 0 | 0 |
| 48A3 | 0 | 0 |
| Average | 1,110 ± 1,324 | 0 ± 0 |
| Transplanted with human CD34+ cells transduced in suspension culture with IL-3, IL-6, SCF, and FL | ||
| 43B1 | 44,900 | 63 |
| 43B2 | 18,100 | 0 |
| 44B1 | 0 | 10 |
| 44B2 | 44,000 | 103 |
| 44B3 | 40,100 | 153 |
| 46C1 | 53,200 | 177 |
| 46C2 | 31,800 | 20 |
| 46C3 | 73,400 | 107 |
| 48B1 | 29,400 | 103 |
| 48B2 | 75,900 | 57 |
| 48B3 | 0 | 0 |
| Average | 37,345 ± 25,299 | 72 ± 62 |
| Transplanted with human CD34+ cells transduced on stromal support with IL-3, IL-6, and SCF | ||
| 43C1 | 73,200 | 90 |
| 43C2 | 110,800 | 223 |
| 43C3 | 34,200 | 7 |
| 44C1 | 78,900 | 240 |
| 44C2 | 28,200 | 213 |
| 46A1 | 99,100 | 326 |
| 46A3 | 52,500 | 170 |
| 48C1 | 40,900 | 203 |
| 48C2 | 1,500 | 0 |
| 48C3 | 32,000 | 233 |
| Average | 55,130 ± 34,493 | 171 ± 106 |
Bone marrow was recovered from bnx mice after engraftment periods of 7 to 8 months. The percentage of human CD45+ cells in the marrow (BM) was determined by FACS, and the number of human cells per 1 × 106 bnx bone marrow cells (total, ungated cells from the FACS profile) was calculated. Human-specific colony-forming assays were plated from the marrow of each mouse to determine the number of clonogenic human hematopoietic progenitors recovered from 1 × 106bnx bone marrow cells.
Between 3 × 105 and 9 × 105 bone marrow cells were plated from each mouse, and numbers were normalized to 1 × 106 for equivalent comparison. Therefore, numbers reported as 0 may be as high as 2 colonies.
Ten thousand events were acquired from each sample, and the numbers were extrapolated to 1 × 106, with a level of sensitivity of 100 human cells per 1 million bnx bone marrow cells.