Maturation Stage of Neutrophilic Cells Grown by SCF + TPO
| . | Differential Counts (%) . | Positive Cells (%) . | ||||||
|---|---|---|---|---|---|---|---|---|
| MB . | PMC . | MC . | MMC . | BC . | Segm . | CD35 . | ALP . | |
| At the beginning of culture | 0.9 | 12.2 | 59.1 | 14.8 | 13.0 | 0 | 0 | 0 |
| SCF + TPO | ||||||||
| 12 hours | 0.8 | 12.3 | 41.5 | 23.1 | 22.3 | 0 | 0 | 0 |
| 24 hours | 0.6 | 1.9 | 29.9 | 36.4 | 30.8 | 0.4 | 0 | 0 |
| 48 hours | 0 | 0.7 | 33.3 | 35.0 | 30.3 | 0.7 | 0 | 0 |
| 72 hours | 0 | 0.7 | 33.8 | 33.8 | 31.0 | 0.7 | 0 | 0 |
| SCF + G-CSF | ||||||||
| 12 hours | 0.6 | 4.3 | 39.0 | 39.0 | 17.1 | 0 | 0 | 0 |
| 24 hours | 0 | 3.2 | 44.2 | 35.9 | 16.0 | 0.7 | 0 | 0 |
| 48 hours | 0 | 0.6 | 30.1 | 34.6 | 30.7 | 4.0 | 4 | 4 |
| 72 hours | 0 | 1.2 | 23.4 | 40.1 | 25.7 | 9.6 | 8 | 9 |
| SCF + GM-CSF | ||||||||
| 12 hours | 0 | 4.6 | 62.1 | 25.6 | 7.7 | 0 | 0 | 0 |
| 24 hours | 0 | 0 | 30.3 | 48.7 | 20.0 | 1.0 | 2 | 0 |
| 48 hours | 0 | 0 | 9.3 | 53.5 | 33.0 | 4.2 | 3 | 0 |
| 72 hours | 0 | 0 | 8.2 | 55.1 | 31.6 | 5.1 | 6 | 0 |
| . | Differential Counts (%) . | Positive Cells (%) . | ||||||
|---|---|---|---|---|---|---|---|---|
| MB . | PMC . | MC . | MMC . | BC . | Segm . | CD35 . | ALP . | |
| At the beginning of culture | 0.9 | 12.2 | 59.1 | 14.8 | 13.0 | 0 | 0 | 0 |
| SCF + TPO | ||||||||
| 12 hours | 0.8 | 12.3 | 41.5 | 23.1 | 22.3 | 0 | 0 | 0 |
| 24 hours | 0.6 | 1.9 | 29.9 | 36.4 | 30.8 | 0.4 | 0 | 0 |
| 48 hours | 0 | 0.7 | 33.3 | 35.0 | 30.3 | 0.7 | 0 | 0 |
| 72 hours | 0 | 0.7 | 33.8 | 33.8 | 31.0 | 0.7 | 0 | 0 |
| SCF + G-CSF | ||||||||
| 12 hours | 0.6 | 4.3 | 39.0 | 39.0 | 17.1 | 0 | 0 | 0 |
| 24 hours | 0 | 3.2 | 44.2 | 35.9 | 16.0 | 0.7 | 0 | 0 |
| 48 hours | 0 | 0.6 | 30.1 | 34.6 | 30.7 | 4.0 | 4 | 4 |
| 72 hours | 0 | 1.2 | 23.4 | 40.1 | 25.7 | 9.6 | 8 | 9 |
| SCF + GM-CSF | ||||||||
| 12 hours | 0 | 4.6 | 62.1 | 25.6 | 7.7 | 0 | 0 | 0 |
| 24 hours | 0 | 0 | 30.3 | 48.7 | 20.0 | 1.0 | 2 | 0 |
| 48 hours | 0 | 0 | 9.3 | 53.5 | 33.0 | 4.2 | 3 | 0 |
| 72 hours | 0 | 0 | 8.2 | 55.1 | 31.6 | 5.1 | 6 | 0 |
CD34−c-kit−/lowCD15+ cells grown by SCF + TPO at 3 weeks were sorted by flow cytometry, and replated at 2,000 cells per well containing SCF + TPO, SCF + G-CSF, or SCF + GM-CSF, as described in Materials and Methods. The cells were serially harvested, and processed for staining with May-Grünwald Giemsa, ALP, or a MoAb for CD35. The results shown are from one representative experiment of three. Similar results were obtained in the other two experiments.
Abbreviations: MB, myeloblasts; PMC, promyelocytes; MC, myelocytes; MMC, metamyelocytes; BC, band cells; Segm, segmented cells; ALP, alkaline phosphatase.