Enrichment of Thymic Reconstitution Activity
| Exp . | Cells Injected . | Thymus Reconstitution* . | Mean No. (Mean %) of DonorDerived Thymocytes . | ||
|---|---|---|---|---|---|
| Cytokines Used . | Phenotype . | Cell No (%) . | |||
| 1 | SF IL-11 | Crude | 6.7 × 105 (100) | 3/3 | 4.6 × 105 (37) |
| Ly-6A/E− | 5.3 × 105 (80) | 0/3 | |||
| Ly-6A/E+ | 8.4 × 104 (12) | 3/3 | 1.1 × 106 (54) | ||
| FcγRII/III−/low | 5.6 × 104 (8.4) | 3/3 | 8.1 × 106 (83) | ||
| FcγRII/IIIhigh | 5.0 × 105 (75) | 0/3 | |||
| 2 | SF IL-11 | Ly-6A/E+ FcγRII/III−/low | 2.8 × 104 (2.3) | 3/3 | 1.8 × 107 (73) |
| Rest of the cells | 1.2 × 106 (95) | 0/3 | |||
| 3 | SF IL-11 IL-7 | Ly-6A/E− | 1.1 × 106 (70) | 0/2 | |
| Ly-6A/E+ | 1.7 × 105 (11) | 1/2 | 5.9 × 104 (3.1) | ||
| FcγRII/III−/low | 1.0 × 105 (6.5) | 2/2 | 1.1 × 105 (8.6) | ||
| FcγRII/IIIhigh | 1.1 × 106 (72) | 0/2 | |||
| 4 | SF IL-11 IL-7 | Ly-6A/E− | 6.0 × 105 (43) | 0/2 | |
| FL | Ly-6A/E+ | 6.0 × 105 (43) | 2/2 | 4.6 × 107 (82) | |
| FcγRII/III−/low | 1.2 × 105 (8.8) | 2/2 | 9.2 × 106 (60) | ||
| FcγRII/IIIhigh | 7.0 × 105 (63) | 0/2 | |||
| Exp . | Cells Injected . | Thymus Reconstitution* . | Mean No. (Mean %) of DonorDerived Thymocytes . | ||
|---|---|---|---|---|---|
| Cytokines Used . | Phenotype . | Cell No (%) . | |||
| 1 | SF IL-11 | Crude | 6.7 × 105 (100) | 3/3 | 4.6 × 105 (37) |
| Ly-6A/E− | 5.3 × 105 (80) | 0/3 | |||
| Ly-6A/E+ | 8.4 × 104 (12) | 3/3 | 1.1 × 106 (54) | ||
| FcγRII/III−/low | 5.6 × 104 (8.4) | 3/3 | 8.1 × 106 (83) | ||
| FcγRII/IIIhigh | 5.0 × 105 (75) | 0/3 | |||
| 2 | SF IL-11 | Ly-6A/E+ FcγRII/III−/low | 2.8 × 104 (2.3) | 3/3 | 1.8 × 107 (73) |
| Rest of the cells | 1.2 × 106 (95) | 0/3 | |||
| 3 | SF IL-11 IL-7 | Ly-6A/E− | 1.1 × 106 (70) | 0/2 | |
| Ly-6A/E+ | 1.7 × 105 (11) | 1/2 | 5.9 × 104 (3.1) | ||
| FcγRII/III−/low | 1.0 × 105 (6.5) | 2/2 | 1.1 × 105 (8.6) | ||
| FcγRII/IIIhigh | 1.1 × 106 (72) | 0/2 | |||
| 4 | SF IL-11 IL-7 | Ly-6A/E− | 6.0 × 105 (43) | 0/2 | |
| FL | Ly-6A/E+ | 6.0 × 105 (43) | 2/2 | 4.6 × 107 (82) | |
| FcγRII/III−/low | 1.2 × 105 (8.8) | 2/2 | 9.2 × 106 (60) | ||
| FcγRII/IIIhigh | 7.0 × 105 (63) | 0/2 | |||
Enriched marrow cells were cultured (40 cells/dish) in methylcellulose with designated cytokines for 10 to 12 days. Approximately 100 primary colonies were pooled and sorted for Ly-6A/E−, Ly-6A/E+, FcγRII/III−/low, and FcγRII/IIIhighcells. In experiment 2, Ly-6A/E+FcγRII/III−/low cells and the rest of the cells were sorted. Crude and sorted cells in proportion to the percentage of the cells in the sorting windows were then injected intravenously into C.B.-17 scid mice. Three weeks after the cell transfer, thymocytes were analyzed for Ly-1.1 expression. Expression of Ly-6A/E and FcγRII/III by pooled primary colony cells and the sorting windows are presented in Fig 3. Reanalysis of the sorted cells are also shown in Fig 3.
Number of reconstituted mice per number of scid mice injected.