EGFP expression in transduced human cord blood cells before and after purification and transplantation into NOD/SCID mice
| Analysis of CB Grafts . | Analysis of NOD/SCID Recipients . | |||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Cord Blood (ref) . | % CD34+ Postinfection . | % EGFP+ Postinfection: . | % EGFP+ Postsorting . | Mouse (ref) . | Graft Size ×106 Cells . | CD45+CD45+/EGFP+CD34+CD34+/EGFP+% EGFP+ . | ||||||||
| Total Cells . | CD34+ . | CD34+/ CD38low . | Total Cells . | in total BM . | CD45+ . | CD34+ . | ||||||||
| CB-2 | 81.1 | 39.2 | 27.2 | 4.7 | (+) | 95.4 | 2I | 0.51 | 10.0 | 6.9 | 1.0 | 4.0 | 69.0 | 40.0 |
| (−) | 0.9 | 2G | 0.29 | 25.6 | 7.5 | 3.8 | 0.7 | 22.4 | 17.5 | |||||
| (−) | 0.9 | 2H | 0.29 | 89.4 | 16.8 | 12.9 | 2.2 | 18.8 | 17.3 | |||||
| CB-3 | 55.2 | 78.8 | 78.6 | N.D. | (+) | 98.1 | 3J | 0.42 | 72.9 | 22.8 | 12.0 | 2.5 | 31.3 | 20.7 |
| (−) | 1.03-151 | 3K3-151 | 0.093-150 | 0.73-151 | NE | NE | NE | NE | NE | |||||
| CB-43-150 | 58.7 | 63.7 | 58.6 | 63.6 | (+) | 96.2 | 4L | 0.35 | 66.9 | 30.9 | 10.7 | 4.5 | 46.3 | 42.3 |
| Mean ± SE | (+) | 96.5 | 0.43 ± 0.05 | 49.9 ± 20.4 | 20.2 ± 7.2 | 7.9 ± 3.5 | 3.7 ± 0.6 | 57.7 ± 11.2 | 41.2 ± 7.0 | |||||
| Mean | (−) | 0.9 | 0.29 | 57.5 | 12.2 | 8.3 | 1.5 | 20.6 | 17.4 | |||||
| Analysis of CB Grafts . | Analysis of NOD/SCID Recipients . | |||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Cord Blood (ref) . | % CD34+ Postinfection . | % EGFP+ Postinfection: . | % EGFP+ Postsorting . | Mouse (ref) . | Graft Size ×106 Cells . | CD45+CD45+/EGFP+CD34+CD34+/EGFP+% EGFP+ . | ||||||||
| Total Cells . | CD34+ . | CD34+/ CD38low . | Total Cells . | in total BM . | CD45+ . | CD34+ . | ||||||||
| CB-2 | 81.1 | 39.2 | 27.2 | 4.7 | (+) | 95.4 | 2I | 0.51 | 10.0 | 6.9 | 1.0 | 4.0 | 69.0 | 40.0 |
| (−) | 0.9 | 2G | 0.29 | 25.6 | 7.5 | 3.8 | 0.7 | 22.4 | 17.5 | |||||
| (−) | 0.9 | 2H | 0.29 | 89.4 | 16.8 | 12.9 | 2.2 | 18.8 | 17.3 | |||||
| CB-3 | 55.2 | 78.8 | 78.6 | N.D. | (+) | 98.1 | 3J | 0.42 | 72.9 | 22.8 | 12.0 | 2.5 | 31.3 | 20.7 |
| (−) | 1.03-151 | 3K3-151 | 0.093-150 | 0.73-151 | NE | NE | NE | NE | NE | |||||
| CB-43-150 | 58.7 | 63.7 | 58.6 | 63.6 | (+) | 96.2 | 4L | 0.35 | 66.9 | 30.9 | 10.7 | 4.5 | 46.3 | 42.3 |
| Mean ± SE | (+) | 96.5 | 0.43 ± 0.05 | 49.9 ± 20.4 | 20.2 ± 7.2 | 7.9 ± 3.5 | 3.7 ± 0.6 | 57.7 ± 11.2 | 41.2 ± 7.0 | |||||
| Mean | (−) | 0.9 | 0.29 | 57.5 | 12.2 | 8.3 | 1.5 | 20.6 | 17.4 | |||||
CD34+ cord blood cells were transduced, sorted on the basis of EGFP expression (see Materials and methods) and transplanted into NOD/SCID mice. Samples were analyzed at the end of the last infection cycle or at 90 to 120 days after transplantation of positive and negative fractions, except for
(40 days).
An aliquot of this CB was also transplanted into mice prior to the cell sorting process (see Tables 1 and 2). (+) positively and (−) negatively selected cells. NE indicates not evaluable; ND indicates not done. CB2 and CB3 samples were stimulated with MGDF/SCF/Flt3-ligand/IL-6. For CB4 stimulation IL-3 was also included.