Table 1.
Immunophenotype of the cells obtained after 6 week LTC-IC-HE of adult human purified CD34−Lin−CD45−CD133− compared with CD34−Lin−CD45−CD133+ cells
| Experiment no. | Purified starting cell population | Cells per well | No. of seeded wells | No. of wells with proliferated cells | No. of wells positive for cells |
||
|---|---|---|---|---|---|---|---|
| CD45+KDR− only | CD45−KDR+ only | CD45+KDR− CD45−KDR+ | |||||
| 1 | CD34−Lin−CD45−CD133− | 1 | 60 | 12 | 8 | 1 | 3 |
| CD34−Lin−CD45−CD133− | 10 | 6 | 6 | 3 | 1 | 2 | |
| 1 | CD34−Lin−CD45−CD133+ | 1 | 60 | 7 | 7 | 0 | 0 |
| CD34−Lin−CD45−CD133+ | 10 | 6 | 6 | 6 | 0 | 0 | |
| 1 | CD34−Lin−CD45+CD133− | 1 | 60 | 14 | 14 | 0 | 0 |
| CD34−Lin−CD45+CD133− | 10 | 6 | 6 | 6 | 0 | 0 | |
| 1 | CD34−Lin−CD45+CD133+ | 1 | 60 | 30 | 30 | 0 | 0 |
| CD34−Lin−CD45+CD133+ | 10 | 6 | 6 | 6 | 0 | 0 | |
Wells from 96-well plates precoated with MS-5 cells were seeded with 1-50 hPB cells of the selected phenotype with the use of the automatic cell device unit of the FACSVantage. Cells were cultured in LTC-IC-HE conditions as described in “Methods.” At the end of the 6-week culture period, individual wells were visually inspected, and wells that contained enough cells were harvested and cells were labeled with lineage-specific Abs. Analysis was performed on the FACSort as described.