Comparison Between Cell Number, CFC, and LTC-IC Injected and Ability to Engraft the BM and the Spleen of NOD/SCID Mice by Human CB Cells Cultured in Stroma-Free Cultures for 4 Weeks
| CB Cells . | Growth Factors Added . | Injected/Mouse . | Engraftment . | ||||||
|---|---|---|---|---|---|---|---|---|---|
| Cells ×106 . | CFC ×103 . | LTC-IC ×103 . | % Human Cells4-151 . | Human CFC/Mouse ×103 ‡ . | |||||
| EXP. 1 | Start of cultures | 0.02 | 2.2 ± 0.7 | 4.62 ± 0.36 | 0.18, 0 | 0.86, 0 | |||
| 4 weeks of cultures | |||||||||
| FL + MGDF | 9.9 ± 2.8 | (495)4-150 | 116.2 ± 18.6 | (52.8)4-150 | 23.42 ± 3.3 | (5.06)4-150 | 10.4, 12 | 20.48 ± 0.4, 86.74 ± 1.3 | |
| SCF + FL + MGDF | 11.7 ± 3.1 | (585)4-150 | 79.3 ± 10.2 | (36)4-150 | 29.97 ± 5.1 | (6.48)4-150 | 11, 15 | ND, 54.71 ± 2.7 | |
| IL6 + SCF + FL + MGDF | 7.4 ± 1.2 | (370)4-150 | 111.6 ± 17.8 | (50.7)4-150 | 23.58 ± 4.2 | (5.12)4-150 | 21, 6 | 25.7 ± 4.6, 46.72 ± 3.7 | |
| EXP. 2 | Start of cultures | 0.02 | 2.46 ± 0.6 | 2.55 ± 0.4 | 0, 0.3 | 0, 1.68 | |||
| 4 weeks of cultures | |||||||||
| FL + MGDF | 4.22 ± 0.77 | (211)4-150 | 48.48 ± 7.1 | (19.7)4-150 | 26.55 ± 4.7 | (11.8)4-150 | 9.8, 0 | 74.26 ± 6.7, 0 | |
| SCF + FL + MGDF | 5.38 ± 0.98 | (269)4-150 | 59.89 ± 8.9 | (24.3)4-150 | 25.82 ± 6.1 | (11.5)4-150 | ND, 21 | ND, 59.64 ± 6.4 | |
| IL6 + SCF + FL + MGDF | 5.6 ± 1.2 | (280)4-150 | 70.72 ± 10.6 | (28.7)4-150 | 30.41 ± 4.6 | (13.5)4-150 | 22.8, 16 | ND, 114.71 ± 11.6 | |
| EXP. 3 | Start of cultures | 0.01 | 0.52 ± 0.5 | 1.7 ± 0.6 | 0, 0 | 0, 0 | |||
| 4 weeks of cultures | |||||||||
| FL + MGDF | 4.16 ± 0.89 | (416)4-150 | 34.24 ± 7.9 | (68.5)4-150 | 38.65 ± 5.1 | (22.7)4-150 | 10.5, 13 | ND, 39.42 ± 5.2 | |
| SCF + FL + MGDF | 6.9 ± 1.5 | (690)4-150 | 43.65 ± 6.9 | (83.9)4-150 | ND | — | 16, 12 | 43.74 ± 5, 110.71 ± 12.6 | |
| IL6 + SCF + FL + MGDF | 7.2 ± 2.2 | (720)4-150 | 42.49 ± 6.3 | (81.7)4-150 | 41.69 ± 8.8 | (24.5)4-150 | 14, 0 | 165.71 ± 13, 0 | |
| CB Cells . | Growth Factors Added . | Injected/Mouse . | Engraftment . | ||||||
|---|---|---|---|---|---|---|---|---|---|
| Cells ×106 . | CFC ×103 . | LTC-IC ×103 . | % Human Cells4-151 . | Human CFC/Mouse ×103 ‡ . | |||||
| EXP. 1 | Start of cultures | 0.02 | 2.2 ± 0.7 | 4.62 ± 0.36 | 0.18, 0 | 0.86, 0 | |||
| 4 weeks of cultures | |||||||||
| FL + MGDF | 9.9 ± 2.8 | (495)4-150 | 116.2 ± 18.6 | (52.8)4-150 | 23.42 ± 3.3 | (5.06)4-150 | 10.4, 12 | 20.48 ± 0.4, 86.74 ± 1.3 | |
| SCF + FL + MGDF | 11.7 ± 3.1 | (585)4-150 | 79.3 ± 10.2 | (36)4-150 | 29.97 ± 5.1 | (6.48)4-150 | 11, 15 | ND, 54.71 ± 2.7 | |
| IL6 + SCF + FL + MGDF | 7.4 ± 1.2 | (370)4-150 | 111.6 ± 17.8 | (50.7)4-150 | 23.58 ± 4.2 | (5.12)4-150 | 21, 6 | 25.7 ± 4.6, 46.72 ± 3.7 | |
| EXP. 2 | Start of cultures | 0.02 | 2.46 ± 0.6 | 2.55 ± 0.4 | 0, 0.3 | 0, 1.68 | |||
| 4 weeks of cultures | |||||||||
| FL + MGDF | 4.22 ± 0.77 | (211)4-150 | 48.48 ± 7.1 | (19.7)4-150 | 26.55 ± 4.7 | (11.8)4-150 | 9.8, 0 | 74.26 ± 6.7, 0 | |
| SCF + FL + MGDF | 5.38 ± 0.98 | (269)4-150 | 59.89 ± 8.9 | (24.3)4-150 | 25.82 ± 6.1 | (11.5)4-150 | ND, 21 | ND, 59.64 ± 6.4 | |
| IL6 + SCF + FL + MGDF | 5.6 ± 1.2 | (280)4-150 | 70.72 ± 10.6 | (28.7)4-150 | 30.41 ± 4.6 | (13.5)4-150 | 22.8, 16 | ND, 114.71 ± 11.6 | |
| EXP. 3 | Start of cultures | 0.01 | 0.52 ± 0.5 | 1.7 ± 0.6 | 0, 0 | 0, 0 | |||
| 4 weeks of cultures | |||||||||
| FL + MGDF | 4.16 ± 0.89 | (416)4-150 | 34.24 ± 7.9 | (68.5)4-150 | 38.65 ± 5.1 | (22.7)4-150 | 10.5, 13 | ND, 39.42 ± 5.2 | |
| SCF + FL + MGDF | 6.9 ± 1.5 | (690)4-150 | 43.65 ± 6.9 | (83.9)4-150 | ND | — | 16, 12 | 43.74 ± 5, 110.71 ± 12.6 | |
| IL6 + SCF + FL + MGDF | 7.2 ± 2.2 | (720)4-150 | 42.49 ± 6.3 | (81.7)4-150 | 41.69 ± 8.8 | (24.5)4-150 | 14, 0 | 165.71 ± 13, 0 | |
CD34+ cells were cultured at 10 × 103/mL or 20 × 103/mL in 24-well plates as described. Cells at the start of cultures and cells harvested after 4 weeks of suspension cultures were cryopreserved and kept frozen until the time of injection in NOD/SCID mice. The expansion cultures were performed in quadruplicate. Two mice were injected each with 20,000 or 10,000 uncultured cells and two mice each with the content of a single expansion culture.
Fold increase (compared with the input value).
Each value represents the percentage of human CD45+, CD71+, and α-GpA+ cells detected in the BM of each individual mouse.
Mean ± SEM of the number of human CFU-GM + BFU-E + CFU-GEMM + CFU-Mk generated by plating 1 × 105 to 5 × 105 BM cells of engrafted NOD/SCID mice (at least three dishes per point), taking into account that BM cells of femurs and tibias represent 25% of the total BM.