Table II.
4 wk
|
8 wk
|
12–18 wk
|
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Cell # | Frequencyreconstitution(#/total) | Average chimerism(range)engraftment type(#/reconstituted) | Frequencyreconstitution(#/total) | Average chimerism(range)engraftment type(#/reconstituted) | Frequencyreconstitution(#/total) | Average chimerism(range)engraftment type(#/reconstituted) |
Untreated BM LT-HSC |
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50 | 100% (8/8) | 11.07% (2.3–22.7%) M + L (8/8) |
100% (7/7) | 29.2% (6.7–56.9%) M + L (7/7) |
100% (7/7) | 30.2% (6.2–56.2%) M + L (7/7) |
D2 Cy/G mobilized BM LT-HSC |
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50 | 46% (7/15) | 2.9% (0.2–12.5%) M + L (7/7) |
40% (6/15) | 6.1% (0.4–26.8%) M + L (4/6), L only (2/6) |
40% (6/15) | 4.6% (0.1–18.1%) M + L (4/6), L only (2/6) |
LT-HSC (Lin−/c-Kit+/Sca-1+/Thy1.1int/Flk-2−) were double FACS–sorted from BM of untreated C57BL/6-Ly5.1 donor mice or from mice subjected to the D2 Cy/G mobilization treatment and tested for their engraftment capacities by limited dilution competitive reconstitution assays. 50 cells were transplanted into lethally irradiated congenic C57BL/6-Ly5.2 recipient mice together with 3 × 105 Ly5.2+ helper BM cells. Recipient mice were considered to be reconstituted by Ly5.1+ donor cells if the frequencies of Ly5.1+ donor-derived myeloid (M, Gr-1+, or Mac-1+) or lymphoid (L, B220+, or CD3+) cells detected in the peripheral blood at the indicated times after transplantation were greater than background levels (≥0.1% as determined by parallel analysis of untransplanted control C57BL/6-Ly5.2 mice). M + L indicates multilineage reconstitution.