HSC frequency and distribution in different subsets of murine lin− fetal liver cells
Phenotype (% of total*) and no. cells per mouse . | No. positive mice/total no. . | HSC frequency†(upper and lower limits) . | % of HSCs in fraction‡ . |
|---|---|---|---|
| CD45+lin− SP (5 ± 2) | 1/18 000 (1/12 000-1/28 000) | 1 | |
| 10 000 | 5/6 | ||
| 8000 | 0/2 | ||
| 2500 | 0/8 | ||
| CD45+ lin− non-SP (14 ± 8) | 1/440 (1/300-1/650) | 99 | |
| 4000 | 7/7 | ||
| 1000 | 6/7 | ||
| 300 | 3/5 | ||
| Lin− Rho− (0.6 ± 0.4) | < 1/7900¶ (< 1/2900-< 1/22 000) | < 1¶ | |
| 1000 | 0/7 | ||
| 480 | 0/3 | ||
| Lin− Rho+ (41 ± 9) | > 1/3300¶ (> 1/2400-> 1/4500) | > 99¶ | |
| 30 000 | 18/18 | ||
| 10 000 | 20/20 |
Phenotype (% of total*) and no. cells per mouse . | No. positive mice/total no. . | HSC frequency†(upper and lower limits) . | % of HSCs in fraction‡ . |
|---|---|---|---|
| CD45+lin− SP (5 ± 2) | 1/18 000 (1/12 000-1/28 000) | 1 | |
| 10 000 | 5/6 | ||
| 8000 | 0/2 | ||
| 2500 | 0/8 | ||
| CD45+ lin− non-SP (14 ± 8) | 1/440 (1/300-1/650) | 99 | |
| 4000 | 7/7 | ||
| 1000 | 6/7 | ||
| 300 | 3/5 | ||
| Lin− Rho− (0.6 ± 0.4) | < 1/7900¶ (< 1/2900-< 1/22 000) | < 1¶ | |
| 1000 | 0/7 | ||
| 480 | 0/3 | ||
| Lin− Rho+ (41 ± 9) | > 1/3300¶ (> 1/2400-> 1/4500) | > 99¶ | |
| 30 000 | 18/18 | ||
| 10 000 | 20/20 |
Values shown represent the mean ± SEM from 4 experiments for the assays of CD45+ lin− SP and non-SP cells, and 3 separate experiments for the assays of lin− Rho− and Rho− cells.
Cells were separated by FACS into the fractions shown and proportionate numbers assayed for multilineage long-term in vivo repopulating ability. HSC frequencies were calculated with data obtained 6 months after transplantation of the SP and non-SP cells, and 4 months after transplantation of the Rho− and Rho+ cells by the method of maximum likelihood using Poisson statistics as described in “Materials and methods.” Values shown in parentheses are the upper and lower limits defined by ± SEM. Levels of PB cells derived from the test transplants in the positive recipients at the time of analysis varied from 4% to 90%.
Calculated as the number of HSCs in each subset expressed as a percentage of the number of HSCs in all the subsets assayed. The number of HSCs in each subset was calculated by multiplying the frequency of HSCs in the subset times the total number of cells in that subset.
Calculated assuming one mouse given the maximum number of Rho− cells tested had been positive, or that one mouse given the minimum number of Rho+ cells had been negative, to indicate the sensitivity of the assays in these instances.