Characteristics of TPO dose-response curves of CD34+CD41+ progenitors
Experiment no. . | Clone size, no. MK . | Clone size, mean NbD (range) . | Plateau no. of clones,*% of total (95% CI) . | TPO50, pg/mL (95% CI) . | CV† . |
|---|---|---|---|---|---|
| 1 | 1 | 0 (0) | 55.1 (51.9-58.3) | 1.26 (0.62-2.55) | 0.62 |
| 1 | 2-3 | 1.16 (1-1.58) | 20.2 (17.3-23.0) | 8.09 (4.87-13.4) | 0.51 |
| 1 | 4-7 | 2.28 (2-2.80) | 18.4 (17.9-19.0) | 41.1 (37.3-45.3) | 0.33 |
| 1 | 8-15 | 3.20 (3-3.90) | 6.23 (6.23-6.23) | 64.7 (64.4-64.9) | 0.19 |
| Total | 1-15 | 0.50 (0-3.90) | 102 (84.0-117)‡ | 4.92 (2.68-9.04) | 1.03 |
| 2 | 1 | 0 (0) | 50.6 (41.6-59.5) | 1.52 (0.64-3.62) | 0.51 |
| 2 | 2-3 | 1.24 (1-1.58) | 19.8 (15.9-23.6) | 6.22 (2.51-15.5) | 0.29 |
| 2 | 4-7 | 2.29 (2-2.80) | 21.8 (19.7-24.0) | 45.7 (34.7-60.2) | 0.31 |
| 2 | 8-15 | 3.21 (3-3.90) | 7.80 (7.80-7.80) | 90.7 (50.0-165) | 0.21 |
| Total | 1-15 | 0.73 (0-3.90) | 96.8 (88.5-105)‡ | 5.50 (3.64-8.00) | 1.00 |
| 3 | 1 | 0 (0) | 43.2 (33.0-53.1) | 2.89 (1.48-5.65) | 0.24 |
| 3 | 2-3 | 1.22 (1-1.58) | 25.6 (21.6-29.6) | 8.95 (7.31-11.0) | 0.26 |
| 3 | 4-7 | 2.30 (2-2.80) | 21.8 (21.1-22.5) | 28.8 (27.7-30.0) | 0.22 |
| 3 | 8-15 | 3.30 (3-3.90) | 9.42 (7.83-11.0) | 120 (88.0-165) | 0.27 |
| Total | 1-15 | 0.83 (0-3.90) | 100 (90.5-110) | 7.31 (5.53-9.66) | 0.47 |
Experiment no. . | Clone size, no. MK . | Clone size, mean NbD (range) . | Plateau no. of clones,*% of total (95% CI) . | TPO50, pg/mL (95% CI) . | CV† . |
|---|---|---|---|---|---|
| 1 | 1 | 0 (0) | 55.1 (51.9-58.3) | 1.26 (0.62-2.55) | 0.62 |
| 1 | 2-3 | 1.16 (1-1.58) | 20.2 (17.3-23.0) | 8.09 (4.87-13.4) | 0.51 |
| 1 | 4-7 | 2.28 (2-2.80) | 18.4 (17.9-19.0) | 41.1 (37.3-45.3) | 0.33 |
| 1 | 8-15 | 3.20 (3-3.90) | 6.23 (6.23-6.23) | 64.7 (64.4-64.9) | 0.19 |
| Total | 1-15 | 0.50 (0-3.90) | 102 (84.0-117)‡ | 4.92 (2.68-9.04) | 1.03 |
| 2 | 1 | 0 (0) | 50.6 (41.6-59.5) | 1.52 (0.64-3.62) | 0.51 |
| 2 | 2-3 | 1.24 (1-1.58) | 19.8 (15.9-23.6) | 6.22 (2.51-15.5) | 0.29 |
| 2 | 4-7 | 2.29 (2-2.80) | 21.8 (19.7-24.0) | 45.7 (34.7-60.2) | 0.31 |
| 2 | 8-15 | 3.21 (3-3.90) | 7.80 (7.80-7.80) | 90.7 (50.0-165) | 0.21 |
| Total | 1-15 | 0.73 (0-3.90) | 96.8 (88.5-105)‡ | 5.50 (3.64-8.00) | 1.00 |
| 3 | 1 | 0 (0) | 43.2 (33.0-53.1) | 2.89 (1.48-5.65) | 0.24 |
| 3 | 2-3 | 1.22 (1-1.58) | 25.6 (21.6-29.6) | 8.95 (7.31-11.0) | 0.26 |
| 3 | 4-7 | 2.30 (2-2.80) | 21.8 (21.1-22.5) | 28.8 (27.7-30.0) | 0.22 |
| 3 | 8-15 | 3.30 (3-3.90) | 9.42 (7.83-11.0) | 120 (88.0-165) | 0.27 |
| Total | 1-15 | 0.83 (0-3.90) | 100 (90.5-110) | 7.31 (5.53-9.66) | 0.47 |
The mean ± 1 SD absolute number of each MK clone size per 96-well Terasaki plate (based on 3 experiments with quadruplicate determinations for each of 8-10 TPO concentrations) was: 1 MK, 26.1 ± 12.6; 2 to 3 MKs, 10.9 ± 3.20; 4 to 7 MKs, 10.4 ± 2.89; 8 to 15 MKs, 3.92 ± 0.63; 1 to 15 MKs, 51.3 ± 20.1. Among 1 MK and 1 to 15 MK clones, the mean absolute number of clones that survived in absence of TPO was 3.83 ± 0.65. In each experiment, four 96-well Terasaki plates were used for each concentration of TPO (total of at least 32 plates).
CV refers to the coefficient of variation (SD/mean) of TPO thresholds.
Total percentage of plateau no. of clones does not equal 100% because the total dose-response curve has been fitted independently of the individual size classes.