Lineage Expression by PKH26-Tracked Donor (Ly-5.1) Cells in Individual Mice
| Cytokines . | PKH26 Staining (total cells) . | Mice . | % B220+ Cells . | % Thy-1.2+ Cells . | % Gr-1+ Mac-1+ Cells . | % Total . | Myeloid/Lymphoid Ratio . |
|---|---|---|---|---|---|---|---|
| TPO | High (3.5 × 104) | 1 | 9.1 | 8.6 | 4.8 | 22.5 | 0.27 |
| 2 | 17.5 | 16.2 | 5.7 | 39.4 | 0.17 | ||
| 3 | 18.8 | 12.3 | 6.9 | 38.0 | 0.22 | ||
| 4 | 23.3 | 21.5 | 4.7 | 49.5 | 0.10 | ||
| Mean ± SD | 37.4 ± 11.1 | 0.19 ± 0.073-150 | |||||
| Low (3.1 × 103) | 1 | 0.9 | |||||
| 2 | 0.3 | ||||||
| 3 | 0.2 | ||||||
| 4 | 0.1 | ||||||
| Mean ± SD | 0.4 ± 0.4 | ||||||
| IL-6, FL | High (4.0 × 105) | 1 | 35.1 | 18.9 | 19.1 | 73.1 | 0.35 |
| 2 | 8.5 | 22.7 | 4.9 | 36.1 | 0.16 | ||
| 3 | 37.3 | 26.2 | 16.0 | 79.5 | 0.25 | ||
| 4 | 30.3 | 13.4 | 32.3 | 76.0 | 0.74 | ||
| 5 | 23.4 | 20.6 | 24.1 | 68.1 | 0.55 | ||
| Mean ± SD | 66.6 ± 17.5 | 0.41 ± 0.233-150 | |||||
| Low (1.2 × 104) | 1 | 6.5 | 2.3 | 1.4 | 10.2 | 0.16 | |
| 2 | 2.2 | 2.2 | 1.7 | 6.1 | 0.39 | ||
| 3 | 2.4 | 1.2 | 1.1 | 4.7 | 0.31 | ||
| 4 | 2.0 | 1.6 | 1.3 | 4.9 | 0.36 | ||
| 5 | 8.2 | 1.1 | 1.1 | 10.4 | 0.12 | ||
| Mean ± SD | 7.3 ± 2.8 | 0.27 ± 0.123-150 |
| Cytokines . | PKH26 Staining (total cells) . | Mice . | % B220+ Cells . | % Thy-1.2+ Cells . | % Gr-1+ Mac-1+ Cells . | % Total . | Myeloid/Lymphoid Ratio . |
|---|---|---|---|---|---|---|---|
| TPO | High (3.5 × 104) | 1 | 9.1 | 8.6 | 4.8 | 22.5 | 0.27 |
| 2 | 17.5 | 16.2 | 5.7 | 39.4 | 0.17 | ||
| 3 | 18.8 | 12.3 | 6.9 | 38.0 | 0.22 | ||
| 4 | 23.3 | 21.5 | 4.7 | 49.5 | 0.10 | ||
| Mean ± SD | 37.4 ± 11.1 | 0.19 ± 0.073-150 | |||||
| Low (3.1 × 103) | 1 | 0.9 | |||||
| 2 | 0.3 | ||||||
| 3 | 0.2 | ||||||
| 4 | 0.1 | ||||||
| Mean ± SD | 0.4 ± 0.4 | ||||||
| IL-6, FL | High (4.0 × 105) | 1 | 35.1 | 18.9 | 19.1 | 73.1 | 0.35 |
| 2 | 8.5 | 22.7 | 4.9 | 36.1 | 0.16 | ||
| 3 | 37.3 | 26.2 | 16.0 | 79.5 | 0.25 | ||
| 4 | 30.3 | 13.4 | 32.3 | 76.0 | 0.74 | ||
| 5 | 23.4 | 20.6 | 24.1 | 68.1 | 0.55 | ||
| Mean ± SD | 66.6 ± 17.5 | 0.41 ± 0.233-150 | |||||
| Low (1.2 × 104) | 1 | 6.5 | 2.3 | 1.4 | 10.2 | 0.16 | |
| 2 | 2.2 | 2.2 | 1.7 | 6.1 | 0.39 | ||
| 3 | 2.4 | 1.2 | 1.1 | 4.7 | 0.31 | ||
| 4 | 2.0 | 1.6 | 1.3 | 4.9 | 0.36 | ||
| 5 | 8.2 | 1.1 | 1.1 | 10.4 | 0.12 | ||
| Mean ± SD | 7.3 ± 2.8 | 0.27 ± 0.123-150 |
Lin−Sca-1+c-kit+ cells (1 × 105) were stained with PKH26 on day 0 and incubated in suspension culture with TPO or with IL-6 and FL for 7 days. PKH26low and PKH26high cells were prepared by sorting and their reconstitution abilities tested. One fourth aliquots of PKH26low (2,720 cells) and PKH26high cells (250 cells) cultured with TPO were injected into each of 4 recipients. One-fifth aliquots of PKH26low(36,415 cells) and PKH26high cells (1,055 cells) cultured with IL-6 and FL were injected into each of 5 recipients. Lineage expression of the engrafted donor cells was analyzed at 5 months posttransplantation. Background (peripheral blood of nontransplanted female mice) staining by anti–Ly-5.1 MoAb (A20-1.7) was less than 1.0%.
There were no significant differences of the myeloid (Gr-1+, Mac-1+ cells)/lymphoid (B220+ cells and Thy-1.2+ cells) ratios among the experimental groups by unpaired t-test.