Expansion of total nucleated cells, CD34+cells, colony-forming cells, and long-term culture-initiating cells under AFT024 noncontact or stroma-free conditions
| Culture condition . | Day 7 fold expansion . | Day 14 fold expansion . | ||||||
|---|---|---|---|---|---|---|---|---|
| TNC . | CD34+ . | CFC . | LTC-IC . | TNC . | CD34+ . | CFC . | LTC-IC . | |
| FS7-SNC* | 10.8 ± 4.1 | 3.2 ± 1.5 | 4.0 ± 0.3 | 2.5 ± 0.8 | 76.1 ± 11.2 | 15.8 ± 10.1 | 1.8 ± 0.3 | 3.0 ± 0.6 |
| FT-SNC* | 10.2 ± 5.8 | 2.2 ± 1.2 | 3.9 ± 0.8 | 4.7 ± 0.8 | 36.6 ± 8.1 | 7.9 ± 4.7 | 2.5 ± 0.3 | 2.4 ± 0.9 |
| Stroma free† | 14.8 ± 3.0 | 1.1 ± 0.2 | 2.2 ± 0.6 | 3.6 ± 1.1 | 185 ± 97 | 2.0 ± 2.0 | 1.0 ± 0.7 | 2.4 ± 0.7 |
| Culture condition . | Day 7 fold expansion . | Day 14 fold expansion . | ||||||
|---|---|---|---|---|---|---|---|---|
| TNC . | CD34+ . | CFC . | LTC-IC . | TNC . | CD34+ . | CFC . | LTC-IC . | |
| FS7-SNC* | 10.8 ± 4.1 | 3.2 ± 1.5 | 4.0 ± 0.3 | 2.5 ± 0.8 | 76.1 ± 11.2 | 15.8 ± 10.1 | 1.8 ± 0.3 | 3.0 ± 0.6 |
| FT-SNC* | 10.2 ± 5.8 | 2.2 ± 1.2 | 3.9 ± 0.8 | 4.7 ± 0.8 | 36.6 ± 8.1 | 7.9 ± 4.7 | 2.5 ± 0.3 | 2.4 ± 0.9 |
| Stroma free† | 14.8 ± 3.0 | 1.1 ± 0.2 | 2.2 ± 0.6 | 3.6 ± 1.1 | 185 ± 97 | 2.0 ± 2.0 | 1.0 ± 0.7 | 2.4 ± 0.7 |
UCB CD34+ cells were cultured in AFT024 noncontact culture or stroma-free culture as described in “Materials and methods.” After 1 and 2 weeks, cells were harvested and analyzed for the number of CD34+ cells, colony-forming cell (CFC), and long-term culture-initiating cell (LTC-IC). Number of CFC in uncultured UCB CD34+ cells = 13.9% ± 5.2%. Number of LTC-IC in uncultured UCB CD34+ cells = 0.6% ± 0.1%. Results are derived from 2 to 3 independent experiments.
AFT024-noncontact culture with FL, SCF, and IL-7 or FL and Tpo (see “Materials and methods” for concentrations).
Stroma-free culture contained granulocyte colony-stimulating factor (G-CSF), macrophage-inflammatory protein-1α (MIP1-α), IL-6, monocyte-chemoattractant protein-1 (MCP-1), vascular endothelial cell growth factor (VEGF), IL-8, N-desulfated, O-sulfated heparin, Tpo, FL, SCF, and IL-7 (see “Materials and methods” for concentrations).
SNC indicates stroma-noncontact. See Table 1 for other abbreviations.