Numbers and erythroid differentiation of clones produced by different subsets of CB34+ CB cells transduced with MIG or p210 and plated in single-cell serum-free liquid cultures
Subsets of CB cells . | Epo . | Total clones (%) . | No. of erythroid clones/no. analyzed (%)* . | ||
---|---|---|---|---|---|
MIG . | p210 . | MIG . | p210 . | ||
CD34+CD45RA−CD71−(stem cell candidates) | + | 69/192 (36) | 103/192 (54) | 8/14 (57) | 20/21 (95) |
− | 24/96 (25) | 41/96 (43) | 8/17 (47) | 12/16 (75) | |
CD34+CD45RA+CD71−(granulopoietic) | + | 62/192 (32) | 84/192 (44) | 7/23 (30) | 21/24 (88) |
− | 17/96 (18) | 22/96 (23) | 1/10 (10) | 8/14 (57) | |
CD34+CD45RA−CD71+(erythroid) | + | 102/192 (53) | 139/192 (72) | 20/22 (91) | 22/22 (100) |
− | 8/96 (8) | 2/96 (2) | 8/8 (100) | 3/3 (100) |
Subsets of CB cells . | Epo . | Total clones (%) . | No. of erythroid clones/no. analyzed (%)* . | ||
---|---|---|---|---|---|
MIG . | p210 . | MIG . | p210 . | ||
CD34+CD45RA−CD71−(stem cell candidates) | + | 69/192 (36) | 103/192 (54) | 8/14 (57) | 20/21 (95) |
− | 24/96 (25) | 41/96 (43) | 8/17 (47) | 12/16 (75) | |
CD34+CD45RA+CD71−(granulopoietic) | + | 62/192 (32) | 84/192 (44) | 7/23 (30) | 21/24 (88) |
− | 17/96 (18) | 22/96 (23) | 1/10 (10) | 8/14 (57) | |
CD34+CD45RA−CD71+(erythroid) | + | 102/192 (53) | 139/192 (72) | 20/22 (91) | 22/22 (100) |
− | 8/96 (8) | 2/96 (2) | 8/8 (100) | 3/3 (100) |
Data shown are the number of clones detected/number of wells analyzed (expressed as a % value in parentheses) for both total clones (≥ 32 cells per well, assessed by direct visualization in situ after 7 days of culture after transduction) and for erythroid clones. The frequency of erythroid clones was determined by morphologic analysis of Wright-Giemsa–stained cytospin preparations made individually on randomly selected 7-day-old clones of ≥ 32 cells/well. Data are from 2 experiments with epo (+) and 1 experiment without epo (−). All wells contained FL, SF, IL-3, IL-6, and G-CSF with or without (±) epo as indicated.