Extent of Human Hematopoietic Cell Content in the Marrow of Long-Term Engrafted bnxMice
| Cell Type and Number . | Duration of Ex Vivo Culture . | Average No. Human Cells/ 1 × 106 bnx BMC3-150 . | Average No. Hu Cells in Total bnx BM3-151 . | Fold Expansion . | Average No. Human CFU per 1 × 106 bnx BM cells3-152 . |
|---|---|---|---|---|---|
| Experiments 1-4 | |||||
| CD34+ 5 × 105 N = 4 | 1 h 3/6/S | 58,675 ± 15,448 | 47.5 ± 12.2 × 105 | 9.5 ± 2.5 | 152 ± 53 |
| CD34+ 5 × 105 N = 10 | 72 h 3/6/S | 26,500 ± 5,773 | 21.2 ± 4.6 × 105 | 3.7 ± 0.8 | 120 ± 27 |
| CD34+38− 2,000 N = 5 | 1 h 3/6/S | 62,380 ± 16,054 | 49.9 ± 12.9 × 105 | 2,495 ± 654 | 279 ± 65 |
| CD34+38− 2,000 N = 14 | 72 h 3/6/S | 32,079 ± 7,581 | 25.7 ± 6.1 × 105 | 1,282 ± 307 | 184 ± 69 |
| Experiments 5 and 6 | |||||
| CD34+38− 2,000 N = 6 | 72 h 3/6/S | 18,500 ± 5,819 | 17.8 ± 4.6 × 105 | 888 ± 227 | 115 ± 43 |
| CD34+38− 2,000 N = 4 | 168 h 3/6/S | 775 ± 484 | 0.6 ± 0.4 × 105 | 31 ± 19 | 8 ± 5 |
| CD34+38− 2,000 N = 6 | 72 h 3/6/S/FL | 31,183 ± 10,151 | 25.0 ± 8.1 × 105 | 1,248 ± 456 | 399 ± 182 |
| CD34+38− 2,000 N = 4 | 168 h 3/6/S/FL | 24,575 ± 10,265 | 19.7 ± 8.2 × 105 | 734 ± 217 | 702 ± 97 |
| Cell Type and Number . | Duration of Ex Vivo Culture . | Average No. Human Cells/ 1 × 106 bnx BMC3-150 . | Average No. Hu Cells in Total bnx BM3-151 . | Fold Expansion . | Average No. Human CFU per 1 × 106 bnx BM cells3-152 . |
|---|---|---|---|---|---|
| Experiments 1-4 | |||||
| CD34+ 5 × 105 N = 4 | 1 h 3/6/S | 58,675 ± 15,448 | 47.5 ± 12.2 × 105 | 9.5 ± 2.5 | 152 ± 53 |
| CD34+ 5 × 105 N = 10 | 72 h 3/6/S | 26,500 ± 5,773 | 21.2 ± 4.6 × 105 | 3.7 ± 0.8 | 120 ± 27 |
| CD34+38− 2,000 N = 5 | 1 h 3/6/S | 62,380 ± 16,054 | 49.9 ± 12.9 × 105 | 2,495 ± 654 | 279 ± 65 |
| CD34+38− 2,000 N = 14 | 72 h 3/6/S | 32,079 ± 7,581 | 25.7 ± 6.1 × 105 | 1,282 ± 307 | 184 ± 69 |
| Experiments 5 and 6 | |||||
| CD34+38− 2,000 N = 6 | 72 h 3/6/S | 18,500 ± 5,819 | 17.8 ± 4.6 × 105 | 888 ± 227 | 115 ± 43 |
| CD34+38− 2,000 N = 4 | 168 h 3/6/S | 775 ± 484 | 0.6 ± 0.4 × 105 | 31 ± 19 | 8 ± 5 |
| CD34+38− 2,000 N = 6 | 72 h 3/6/S/FL | 31,183 ± 10,151 | 25.0 ± 8.1 × 105 | 1,248 ± 456 | 399 ± 182 |
| CD34+38− 2,000 N = 4 | 168 h 3/6/S/FL | 24,575 ± 10,265 | 19.7 ± 8.2 × 105 | 734 ± 217 | 702 ± 97 |
Bone marrow was recovered from bnx mice and the percentage of human CD45+ cells in the marrow (BM) was determined by FACS. The number of human cells per 1 × 106bnx bone marrow cells was calculated. Human-specific colony-forming assays were plated from the marrow of each mouse to determine the number of clonogenic human progenitors recovered from 1 × 106bnxmarrow cells.
The limits of detection by FACS analysis were 100 human hematopoietic (CD45+) cells per 1 × 106bnxbone marrow cells.
Assuming an average of 80 × 106 total bone marrow cells per mouse and 100% seeding efficiency for transplanted cells.
Between 3 × 105 and 9 × 105 bone marrow cells were plated from each mouse, and numbers were normalized to 1 × 106. Therefore, numbers reported as 0 may be as high as two colonies.