IL-6 alters the lineage decision of hematopoietic stem cells
Culture and genotype . | IL-6 added . | Primitive, % wells . | Myeloid, % wells . | B lymphoid, % wells . | Myeloid + B lymphoid, % wells . | No. positive colonies/no. plated wells . |
|---|---|---|---|---|---|---|
| OP9 culture | ||||||
| Wild-type; RAG1-GFP | No | 6.9 | 31.0 | 20.7 | 41.4 | 29/120 |
| Wild-type; RAG1-GFP | Yes | 15.4 | 76.9 | 2.6 | 5.1 | 39/120 |
| SHIP-/-; RAG1-GFP | No | 4.3 | 52.2 | 17.4 | 26.1 | 23/120 |
| SHIP-/-; RAG1-GFP | Yes | 7.1 | 92.9 | 0.0 | 0.0 | 28/120 |
| Serum-free, stroma-free culture | ||||||
| Wild-type; RAG1-GFP | No | 0 | 20.0 | 20.0 | 40.0 | 5/240* |
| Wild-type; RAG1-GFP | Yes | 5.7 | 94.3 | 0.0 | 0.0 | 35/240 |
| SHIP-/-; RAG1-GFP | No | 22.2 | 77.8 | 0.0 | 0.0 | 18/120 |
| SHIP-/-; RAG1-GFP | Yes | 11.4 | 88.6 | 0.0 | 0.0 | 44/120 |
Culture and genotype . | IL-6 added . | Primitive, % wells . | Myeloid, % wells . | B lymphoid, % wells . | Myeloid + B lymphoid, % wells . | No. positive colonies/no. plated wells . |
|---|---|---|---|---|---|---|
| OP9 culture | ||||||
| Wild-type; RAG1-GFP | No | 6.9 | 31.0 | 20.7 | 41.4 | 29/120 |
| Wild-type; RAG1-GFP | Yes | 15.4 | 76.9 | 2.6 | 5.1 | 39/120 |
| SHIP-/-; RAG1-GFP | No | 4.3 | 52.2 | 17.4 | 26.1 | 23/120 |
| SHIP-/-; RAG1-GFP | Yes | 7.1 | 92.9 | 0.0 | 0.0 | 28/120 |
| Serum-free, stroma-free culture | ||||||
| Wild-type; RAG1-GFP | No | 0 | 20.0 | 20.0 | 40.0 | 5/240* |
| Wild-type; RAG1-GFP | Yes | 5.7 | 94.3 | 0.0 | 0.0 | 35/240 |
| SHIP-/-; RAG1-GFP | No | 22.2 | 77.8 | 0.0 | 0.0 | 18/120 |
| SHIP-/-; RAG1-GFP | Yes | 11.4 | 88.6 | 0.0 | 0.0 | 44/120 |
Sorted Lin-c-kithiSca-1+GFP- progenitors were cultured with OP9 stromal cells or in serum-free, stromal-free conditions in media containing cytokines (SCF, 20 ng/mL; FL, 100 ng/mL; IL-7, 2 ng/mL). IL-6 was added to 2 ng/mL as indicated. After 10 days (OP9 cultures) or 14 days (cytokine cultures), the individual wells were analyzed for CD19, Mac-1, and GFP by flow cytometry. The identity of the clone (primitive, myeloid, B lymphoid, or mixed) was determined as shown in Figure 4. Shown are the percentage of wells having the indicated phenotype. The experiment is representative of 2 identical trials.
One of the 5 colonies had a phenotype that could not be determined.