Replating Studies of Blast Colonies Derived From Clone-Sorted Lin−c-Kit+Sca-1+Cells
| Mice . | Colony No. . | Cell No. Per Colony . | Stimuli in Primary Culture . | No. of Secondary Colonies Per ½ Primary Colony . | Replating Efficiency (%) . | ||||||
|---|---|---|---|---|---|---|---|---|---|---|---|
| GM . | B . | Mk . | Mast . | Mix . | Blast . | Total . | |||||
| Tg mice | 1 | 110 | hG-CSF | 31 | 3 | 2 | 2 | 13 | 1 | 52 | 94 |
| 2 | 210 | 53 | 4 | 1 | 1 | 27 | 0 | 86 | 82 | ||
| 3 | 83 | 29 | 1 | 1 | 0 | 10 | 1 | 42 | 100 | ||
| 4 | 156 | 39 | 2 | 1 | 2 | 11 | 1 | 56 | 72 | ||
| 5 | 134 | 41 | 2 | 2 | 1 | 17 | 0 | 63 | 94 | ||
| 6 | 230 | 80 | 3 | 1 | 2 | 21 | 2 | 109 | 95 | ||
| 7 | 130 | 35 | 3 | 2 | 3 | 14 | 1 | 58 | 89 | ||
| 8 | 168 | 52 | 4 | 2 | 1 | 19 | 1 | 79 | 94 | ||
| 9 | 96 | 25 | 3 | 2 | 1 | 13 | 1 | 45 | 93 | ||
| 10 | 124 | 44 | 2 | 1 | 3 | 12 | 0 | 62 | 102 | ||
| 11 | 96 | mIL-3 | 36 | 1 | 1 | 3 | 9 | 0 | 48 | 100 | |
| 12 | 81 | 27 | 2 | 1 | 0 | 6 | 1 | 37 | 92 | ||
| 13 | 64 | 18 | 1 | 1 | 1 | 7 | 1 | 29 | 90 | ||
| 14 | 95 | 27 | 2 | 1 | 2 | 10 | 1 | 43 | 89 | ||
| 15 | 176 | 53 | 3 | 2 | 2 | 26 | 0 | 86 | 98 | ||
| Littermates | 16 | 124 | mIL-3 | 45 | 3 | 1 | 1 | 13 | 0 | 63 | 102 |
| 17 | 74 | 15 | 1 | 1 | 0 | 8 | 1 | 26 | 71 | ||
| 18 | 84 | 28 | 1 | 2 | 1 | 9 | 1 | 41 | 98 | ||
| 19 | 192 | 68 | 3 | 1 | 2 | 24 | 1 | 99 | 103 | ||
| 20 | 102 | 33 | 2 | 1 | 2 | 12 | 1 | 50 | 99 | ||
| Mice . | Colony No. . | Cell No. Per Colony . | Stimuli in Primary Culture . | No. of Secondary Colonies Per ½ Primary Colony . | Replating Efficiency (%) . | ||||||
|---|---|---|---|---|---|---|---|---|---|---|---|
| GM . | B . | Mk . | Mast . | Mix . | Blast . | Total . | |||||
| Tg mice | 1 | 110 | hG-CSF | 31 | 3 | 2 | 2 | 13 | 1 | 52 | 94 |
| 2 | 210 | 53 | 4 | 1 | 1 | 27 | 0 | 86 | 82 | ||
| 3 | 83 | 29 | 1 | 1 | 0 | 10 | 1 | 42 | 100 | ||
| 4 | 156 | 39 | 2 | 1 | 2 | 11 | 1 | 56 | 72 | ||
| 5 | 134 | 41 | 2 | 2 | 1 | 17 | 0 | 63 | 94 | ||
| 6 | 230 | 80 | 3 | 1 | 2 | 21 | 2 | 109 | 95 | ||
| 7 | 130 | 35 | 3 | 2 | 3 | 14 | 1 | 58 | 89 | ||
| 8 | 168 | 52 | 4 | 2 | 1 | 19 | 1 | 79 | 94 | ||
| 9 | 96 | 25 | 3 | 2 | 1 | 13 | 1 | 45 | 93 | ||
| 10 | 124 | 44 | 2 | 1 | 3 | 12 | 0 | 62 | 102 | ||
| 11 | 96 | mIL-3 | 36 | 1 | 1 | 3 | 9 | 0 | 48 | 100 | |
| 12 | 81 | 27 | 2 | 1 | 0 | 6 | 1 | 37 | 92 | ||
| 13 | 64 | 18 | 1 | 1 | 1 | 7 | 1 | 29 | 90 | ||
| 14 | 95 | 27 | 2 | 1 | 2 | 10 | 1 | 43 | 89 | ||
| 15 | 176 | 53 | 3 | 2 | 2 | 26 | 0 | 86 | 98 | ||
| Littermates | 16 | 124 | mIL-3 | 45 | 3 | 1 | 1 | 13 | 0 | 63 | 102 |
| 17 | 74 | 15 | 1 | 1 | 0 | 8 | 1 | 26 | 71 | ||
| 18 | 84 | 28 | 1 | 2 | 1 | 9 | 1 | 41 | 98 | ||
| 19 | 192 | 68 | 3 | 1 | 2 | 24 | 1 | 99 | 103 | ||
| 20 | 102 | 33 | 2 | 1 | 2 | 12 | 1 | 50 | 99 | ||
Single-cell cultures of Lin−c-Kit+Sca-1+ BM cells from hG-CSFR Tg mice and their littermates were incubated with hG-CSF (20 ng/mL) or mIL-3 (10 ng/mL). On day 8 of incubation, blast colonies were individually lifted from the culture and replated to a secondary culture containing mSCF (100 ng/mL), mIL-3 (10 ng/mL), EPO (2 U/mL), TPO (20 ng/mL), and IL-6 (100 ng/mL). Data represent the number of colonies on day 8 of the secondary culture.