Stroma-Contact or Stroma-Noncontact Cocultures of Human CB CD34+ Cells With AGM-S3 Cells
| Duration of Coculture . | Culture Conditions . | No. of Progenitors/Well . | |||
|---|---|---|---|---|---|
| CFU-GM . | BFU-E . | CFU-Mix . | Total . | ||
| 0 | 56 ± 6 | 26 ± 4 | 54 ± 12 | 136 ± 16 | |
| 2 wk | Stroma-free | 8 ± 3 | 0 | 0 | 8 ± 3 |
| Noncontact | 96 ± 16 | 0 | 0 | 96 ± 16 | |
| Contact | 248 ± 36 | 180 ± 28 | 108 ± 20 | 536 ± 56 | |
| 4 wk | Stroma-free | 0 | 0 | 0 | 0 |
| Noncontact | 64 ± 12 | 0 | 0 | 64 ± 12 | |
| Contact | 430 ± 40 | 80 ± 20 | 60 ± 10 | 430 ± 40 | |
| Duration of Coculture . | Culture Conditions . | No. of Progenitors/Well . | |||
|---|---|---|---|---|---|
| CFU-GM . | BFU-E . | CFU-Mix . | Total . | ||
| 0 | 56 ± 6 | 26 ± 4 | 54 ± 12 | 136 ± 16 | |
| 2 wk | Stroma-free | 8 ± 3 | 0 | 0 | 8 ± 3 |
| Noncontact | 96 ± 16 | 0 | 0 | 96 ± 16 | |
| Contact | 248 ± 36 | 180 ± 28 | 108 ± 20 | 536 ± 56 | |
| 4 wk | Stroma-free | 0 | 0 | 0 | 0 |
| Noncontact | 64 ± 12 | 0 | 0 | 64 ± 12 | |
| Contact | 430 ± 40 | 80 ± 20 | 60 ± 10 | 430 ± 40 | |
AGM-S3 cell layers were prepared in 24-well plates. Five hundred human CB CD34+ cells per well were planted on the stromal cell layers (contact), in transwell inserts placed above the stromal cell layers (noncontact), or in transwell inserts placed in empty wells (stroma-free). Clonogenic progenitors generated in the culture at each time point were evaluated by culturing the total or a fraction of the cells harvested. The colonies were counted at day 14 of culture. The number of colonies indicates the mean ± SD of triplicate cultures.