Limited dilution analysis of human FL engraftment in NOD/SCID mice
FL specimen and cell dose . | No. engrafted mice . | No. mice receiving transplants . |
|---|---|---|
| FL no. 841 | ||
| Unexpanded, d 0* | ||
| 125 000 | 2 | 4 |
| 250 000 | 1 | 5 |
| 500 000 | 2 | 4 |
| 1 000 000 | 5 | 5 |
| 2 500 000 | 4 | 5 |
| 5 000 000 | 4 | 4 |
| 10 000 000 | 1 | 1 |
| 20 000 000 | 2 | 2 |
| Expanded, d 28† | ||
| 250 000 | 1 | 5 |
| 500 000 | 4 | 5 |
| 1 000 000 | 8 | 8 |
| 2 500 000 | 4 | 4 |
| 5 000 000 | 5 | 5 |
| 10 000 000 | 7 | 7 |
| FL no. 852 | ||
| Unexpanded, d 0‡ | ||
| 62 500 | 0 | 3 |
| 125 000 | 0 | 5 |
| 250 000 | 1 | 4 |
| 500 000 | 1 | 5 |
| 1 000000 | 1 | 4 |
| 5 000 000 | 3 | 4 |
| Expanded, d 28§ | ||
| 250 000 | 0 | 3 |
| 500 000 | 5 | 6 |
| 1 000 000 | 7 | 9 |
| 2 500 000 | 5 | 5 |
| 5 000 000 | 3 | 3 |
| 10 000 000 | 4 | 4 |
| FL no. 873 | ||
| Unexpanded, d 0∥ | ||
| 62 500 | 1 | 3 |
| 125 000 | 1 | 4 |
| 250 000 | 1 | 2 |
| 500 000 | 3 | 3 |
| 1 000 000 | 5 | 6 |
| 3 000 000 | 3 | 3 |
| 10 000 000 | 3 | 3 |
| Expanded, d 28¶ | ||
| 250 000 | 0 | 5 |
| 500 000 | 0 | 5 |
| 1 000 000 | 4 | 10 |
| 2 500 000 | 4 | 5 |
| 5 000 000 | 4 | 4 |
| 10 000 000 | 5 | 5 |
FL specimen and cell dose . | No. engrafted mice . | No. mice receiving transplants . |
|---|---|---|
| FL no. 841 | ||
| Unexpanded, d 0* | ||
| 125 000 | 2 | 4 |
| 250 000 | 1 | 5 |
| 500 000 | 2 | 4 |
| 1 000 000 | 5 | 5 |
| 2 500 000 | 4 | 5 |
| 5 000 000 | 4 | 4 |
| 10 000 000 | 1 | 1 |
| 20 000 000 | 2 | 2 |
| Expanded, d 28† | ||
| 250 000 | 1 | 5 |
| 500 000 | 4 | 5 |
| 1 000 000 | 8 | 8 |
| 2 500 000 | 4 | 4 |
| 5 000 000 | 5 | 5 |
| 10 000 000 | 7 | 7 |
| FL no. 852 | ||
| Unexpanded, d 0‡ | ||
| 62 500 | 0 | 3 |
| 125 000 | 0 | 5 |
| 250 000 | 1 | 4 |
| 500 000 | 1 | 5 |
| 1 000000 | 1 | 4 |
| 5 000 000 | 3 | 4 |
| Expanded, d 28§ | ||
| 250 000 | 0 | 3 |
| 500 000 | 5 | 6 |
| 1 000 000 | 7 | 9 |
| 2 500 000 | 5 | 5 |
| 5 000 000 | 3 | 3 |
| 10 000 000 | 4 | 4 |
| FL no. 873 | ||
| Unexpanded, d 0∥ | ||
| 62 500 | 1 | 3 |
| 125 000 | 1 | 4 |
| 250 000 | 1 | 2 |
| 500 000 | 3 | 3 |
| 1 000 000 | 5 | 6 |
| 3 000 000 | 3 | 3 |
| 10 000 000 | 3 | 3 |
| Expanded, d 28¶ | ||
| 250 000 | 0 | 5 |
| 500 000 | 0 | 5 |
| 1 000 000 | 4 | 10 |
| 2 500 000 | 4 | 5 |
| 5 000 000 | 4 | 4 |
| 10 000 000 | 5 | 5 |
NOD/SCID mice were given transplants with serial doses of total nucleated cells from 3 different human FL samples before or following 28 days of culture. Murine BM was analyzed 8 weeks after transplantation by flow cytometry and confirmed by Southern blot hybridization. Mice were scored as engrafted if a specific band was detectable by Southern blot analysis, as shown in Figure 1D (limit of detection, about 0.1% human cells). Poisson statistics were applied to the single-hit model, and the frequency of SRC calculated with the maximum likelihood estimator.
SRC frequency: 1 in 704 503 nucleated cells; 95% confidence limits: 1 in 370 751 to 1 in 1 338 698; (approximately 7.0 % CD45, or approximately 1 SRC in 49 000 CD45+ cells).
SRC frequency: 1 in 365 149 nucleated cells; 95% confidence limits: 1 in 196 036 to 1 in 680 151. Approximate CD45+ cell expansion: 343-fold; net SRC expansion: about 46-fold.
SRC frequency: 1 in 2 982 936 nucleated cells; 95% confidence limits: 1 in 1 249 523 to 1 in 7 121 047 (approximately 8.7% CD45, or approximately 1 SRC in 260 000 CD45+ cells).
SRC frequency: 1 in 556 329 nucleated cells; 95% confidence limits: 1 in 310 878 to 1 in 995 575. Approximate CD45+ cell expansion: 402-fold; net SRC expansion: about 188-fold.
SRC frequency: 1 in 366 045 nucleated cells; 95% confidence limits: 1 in 180 564 to 1 in 742 055 (approximately 11.8% CD45, or approximately 1 SRC in 43 000 CD45+ cells).
SRC frequency: 1 in 1 994 478 nucleated cells; 95% confidence limits: 1 in 1 126 292 to 1 in 3 531 893. Approximate CD45+ cell expansion: 449-fold; net SRC expansion: about 10-fold.