Determination of the Cell Number, CFC, LTC-IC, and the Ability to Engraft the BM of NOD/SCID Recipients by Human CB Cells During Stroma-Free Cultures
| In Culture . | Injected/Mouse . | Engraftment . | |||||||
|---|---|---|---|---|---|---|---|---|---|
| Source of Cells . | Growth Factors Added . | Cells (fold increase) . | CFC (fold increase) . | LTC-IC (fold increase) . | Cells . | CFC . | LTC-IC . | % of Human Cells5-150 . | Human CFC/ mouse ×103 5-151 . |
| Start | 10,000 | 866.4 | 1,332 | 10,000 | 866.4 | 1,332 | 0, 0 | 0, 0 | |
| 5,000 | 433 | 666 | 0, 0 | 0, 0 | |||||
| 2,500 | 216 | 333 | 0, 0 | 0, 0 | |||||
| 1,250 | 108 | 166 | 0, 0 | 0, 0 | |||||
| 4 weeks | +IL-6 + SCF + FL + MGDF | 3.16 × 106 (316) | 37,184 (43) | 11,248 (8.4) | 3.160 × 106 0.395 × 106 | 37,184 4,648 | 11,248 1,456 | 8.5, 9.9 2.5, 3 | 15.64, 18.4 7.2, 9.6 |
| 8 weeks | +IL-6 + SCF + FL + MGDF | 28.99 × 106 (2,899) | 129,464 (149) | 56,304 (42) | 28.99 × 106 3.60 × 106 | 129,464 16,183 | 56,304 7,038 | 7.3, 18 2.8, 3.6 | 18.22, 36.72 ND, 34.32 |
| 1.80 × 106 | 8,091 | 3,519 | 0.7, 1.9 | 1.34, 3.87 | |||||
| 12 weeks | +IL-6 + SCF + FL + MGDF | 109 × 106 (10,900) | 2,909,061 (3,358) | 382,284 (287) | 109 × 106 13.6 × 106 | — 363,632 | — 47,785 | — — 11, 16 | 34.98, 50.76 |
| 6.8 × 106 | 181,816 | 23,892 | 8, 6.8 | 23.44, 44.66 | |||||
| 3.4 × 106 | 90,908 | 11,946 | 1.4, 0.9 | 12.6, 19.3 | |||||
| Start | 20,000 | 2,248 | 3,632 | 20,000 | 3,248 | 3,632 | 0, 0 | 2.08, 0 | |
| 5,000 | 812 | 908 | 0, 0 | 0, 0 | |||||
| 2,500 | 406 | 454 | 0, 0 | 0, 0 | |||||
| 1,250 | 203 | 227 | 0, 0 | 0, 0 | |||||
| 7 weeks | FL + MGDF | 22 × 106 (1,100) | 372,736 (166) | 90,112 (25) | 22 × 106 2.75 × 106 | 124,930 15,616 | 90,112 11,264 | 6.8, 15 4, 11 | 29.9, 63.84 8.3, 75.9 |
| 1.37 × 106 | 7,808 | 5,632 | 0, 2.6 | 0, 13.4 | |||||
| SCF + FL + MGDF | 36.4 × 106 (1,820) | 381,728 (170) | 126,976 (35) | 36.40 × 106 4.55 × 106 | 381,728 47,716 | 126,976 15,872 | 7.8, 14.2 5.6, 9 | 41.87, 46.86 2.08, ND | |
| 2.27 × 106 | 23,858 | 7,936 | 0.9, 3 | 2.54, 10.1 | |||||
| IL-6 + SCF + FL + MGDF | 17.8 × 106 (890) | 361,081 (160) | 102,400 (28) | 17.80 × 106 2.22 × 106 | 361,081 45,135 | 102,400 12,800 | 18, 10.4 4, 6 | 108.9, 54.16 ND, 23.9 | |
| 1.11 × 106 | 22,567 | 6,400 | 1, 2.1 | 6.32, 15.6 | |||||
| 12 weeks | IL-6 + SCF + FL + MGDF | 710 × 106 (35,500) | 2,928,640 (1,303) | 980,460 (270) | — 88.75 × 106 | — 366,080 | — 122,580 | — — ND, ND | — — ND, ND |
| 22.20 × 106 | 91,520 | 30,641 | 11, 6.5 | 50.16, 30.07 | |||||
| 11.10 × 106 | 45,760 | 15,322 | 0.6, 3.9 | 1.87, 6 | |||||
| 5.50 × 106 | 22,880 | 7,661 | 0.9, 0.5 | 5.56, 2.3 | |||||
| In Culture . | Injected/Mouse . | Engraftment . | |||||||
|---|---|---|---|---|---|---|---|---|---|
| Source of Cells . | Growth Factors Added . | Cells (fold increase) . | CFC (fold increase) . | LTC-IC (fold increase) . | Cells . | CFC . | LTC-IC . | % of Human Cells5-150 . | Human CFC/ mouse ×103 5-151 . |
| Start | 10,000 | 866.4 | 1,332 | 10,000 | 866.4 | 1,332 | 0, 0 | 0, 0 | |
| 5,000 | 433 | 666 | 0, 0 | 0, 0 | |||||
| 2,500 | 216 | 333 | 0, 0 | 0, 0 | |||||
| 1,250 | 108 | 166 | 0, 0 | 0, 0 | |||||
| 4 weeks | +IL-6 + SCF + FL + MGDF | 3.16 × 106 (316) | 37,184 (43) | 11,248 (8.4) | 3.160 × 106 0.395 × 106 | 37,184 4,648 | 11,248 1,456 | 8.5, 9.9 2.5, 3 | 15.64, 18.4 7.2, 9.6 |
| 8 weeks | +IL-6 + SCF + FL + MGDF | 28.99 × 106 (2,899) | 129,464 (149) | 56,304 (42) | 28.99 × 106 3.60 × 106 | 129,464 16,183 | 56,304 7,038 | 7.3, 18 2.8, 3.6 | 18.22, 36.72 ND, 34.32 |
| 1.80 × 106 | 8,091 | 3,519 | 0.7, 1.9 | 1.34, 3.87 | |||||
| 12 weeks | +IL-6 + SCF + FL + MGDF | 109 × 106 (10,900) | 2,909,061 (3,358) | 382,284 (287) | 109 × 106 13.6 × 106 | — 363,632 | — 47,785 | — — 11, 16 | 34.98, 50.76 |
| 6.8 × 106 | 181,816 | 23,892 | 8, 6.8 | 23.44, 44.66 | |||||
| 3.4 × 106 | 90,908 | 11,946 | 1.4, 0.9 | 12.6, 19.3 | |||||
| Start | 20,000 | 2,248 | 3,632 | 20,000 | 3,248 | 3,632 | 0, 0 | 2.08, 0 | |
| 5,000 | 812 | 908 | 0, 0 | 0, 0 | |||||
| 2,500 | 406 | 454 | 0, 0 | 0, 0 | |||||
| 1,250 | 203 | 227 | 0, 0 | 0, 0 | |||||
| 7 weeks | FL + MGDF | 22 × 106 (1,100) | 372,736 (166) | 90,112 (25) | 22 × 106 2.75 × 106 | 124,930 15,616 | 90,112 11,264 | 6.8, 15 4, 11 | 29.9, 63.84 8.3, 75.9 |
| 1.37 × 106 | 7,808 | 5,632 | 0, 2.6 | 0, 13.4 | |||||
| SCF + FL + MGDF | 36.4 × 106 (1,820) | 381,728 (170) | 126,976 (35) | 36.40 × 106 4.55 × 106 | 381,728 47,716 | 126,976 15,872 | 7.8, 14.2 5.6, 9 | 41.87, 46.86 2.08, ND | |
| 2.27 × 106 | 23,858 | 7,936 | 0.9, 3 | 2.54, 10.1 | |||||
| IL-6 + SCF + FL + MGDF | 17.8 × 106 (890) | 361,081 (160) | 102,400 (28) | 17.80 × 106 2.22 × 106 | 361,081 45,135 | 102,400 12,800 | 18, 10.4 4, 6 | 108.9, 54.16 ND, 23.9 | |
| 1.11 × 106 | 22,567 | 6,400 | 1, 2.1 | 6.32, 15.6 | |||||
| 12 weeks | IL-6 + SCF + FL + MGDF | 710 × 106 (35,500) | 2,928,640 (1,303) | 980,460 (270) | — 88.75 × 106 | — 366,080 | — 122,580 | — — ND, ND | — — ND, ND |
| 22.20 × 106 | 91,520 | 30,641 | 11, 6.5 | 50.16, 30.07 | |||||
| 11.10 × 106 | 45,760 | 15,322 | 0.6, 3.9 | 1.87, 6 | |||||
| 5.50 × 106 | 22,880 | 7,661 | 0.9, 0.5 | 5.56, 2.3 | |||||
The value in parentheses represents the fold increase compared with input values.
Each value represents the percent of human CD45+, CD71+, and α-GPA+ cells in the BM of each individual mouse.
As explained in Table 4. Expansion studies were performed in quadruplicate. Two mice were injected with each cell dose as indicated.