In vitro cycling SRCs up-regulate Fas expression
Experiment, expansion, and sorted cells . | % of total Lin− cells . | Cells transplanted per mouse, no. . | Positive mice/total mice, no. . | Human engraftment, % (replicate %) . |
|---|---|---|---|---|
| Exp 1, 15-fold expansion | ||||
| CB Lin−Fas− | 42 | 450 000 | 0/2 | 0 (0-0) |
| CB Lin−Fas+ | 58 | 450 000 | 4/4 | 3 (0.3-0.3-0.4-10) |
| Exp 2, 35-fold expansion | ||||
| CB Lin−Fas− | 38 | 1.8 × 106 | 1/6 | 0.3 (0-0-0-0-0-0-2) |
| CB Lin−Fas+ | 62 | 1.8 × 106 | 7/7 | 7 (0.3-5-5-6-7-10-15) |
| Exp 3, 30-fold expansion | ||||
| CB Lin−Fas− | 40 | 1.5 × 106 | 1/6 | 0.05 (0-0-0-0-0-0.3) |
| CB Lin−Fas+ | 60 | 1.5 × 106 | 5/5 | 2.4 (0.4-0.7-0.7-3-7) |
| Exp 4, 19-fold expansion | ||||
| CB Lin− | 100 | 950 000 | 3/3 | 13 (10-11-18) |
| CB Lin−Fas+ | 70 | 950 000 | 4/4 | 28 (22-23-26-40) |
Experiment, expansion, and sorted cells . | % of total Lin− cells . | Cells transplanted per mouse, no. . | Positive mice/total mice, no. . | Human engraftment, % (replicate %) . |
|---|---|---|---|---|
| Exp 1, 15-fold expansion | ||||
| CB Lin−Fas− | 42 | 450 000 | 0/2 | 0 (0-0) |
| CB Lin−Fas+ | 58 | 450 000 | 4/4 | 3 (0.3-0.3-0.4-10) |
| Exp 2, 35-fold expansion | ||||
| CB Lin−Fas− | 38 | 1.8 × 106 | 1/6 | 0.3 (0-0-0-0-0-0-2) |
| CB Lin−Fas+ | 62 | 1.8 × 106 | 7/7 | 7 (0.3-5-5-6-7-10-15) |
| Exp 3, 30-fold expansion | ||||
| CB Lin−Fas− | 40 | 1.5 × 106 | 1/6 | 0.05 (0-0-0-0-0-0.3) |
| CB Lin−Fas+ | 60 | 1.5 × 106 | 5/5 | 2.4 (0.4-0.7-0.7-3-7) |
| Exp 4, 19-fold expansion | ||||
| CB Lin− | 100 | 950 000 | 3/3 | 13 (10-11-18) |
| CB Lin−Fas+ | 70 | 950 000 | 4/4 | 28 (22-23-26-40) |
CD34+ CB cells were cultured in SF medium supplemented with SFT3 for 5 days, at which time Lin−Fas− (or total Lin− cells; experiment 4) and Lin−Fas+ cells were sorted on a FACSVantage. The expansion equivalents (EEs) of 30 000 to 50 000 CD34+ CB cells were transplanted per mouse. Data shown for human engraftment in NOD-SCID mice are mean values of individual mice per group, with replicate percentages shown in parentheses. Positive mice were defined as mice with human reconstitution of both myeloid and B-cell lineages (“Materials and methods”).
SRC indicates SCID-repopulating cell; exp, experiment.