Fig. 1.
Fig. 1. Expression of c-kit on long-term reconstituting stem cells from normal and 5-FU–treated bone marrow. Erythrocyte-depleted WBM from normal mice was stained with anti–c-kit and the lineage cocktail and was sorted into 3 populations as indicated (A, WBM). The relative representation of each population is shown as a percentage within each gate. For comparison, the relative representation of the 3 populations in lineage− bone marrow is shown (B, lin−). WBM cells were sorted based only on the expression of c-kit into a c-kit− population (79% of bone marrow), c-kitlow population (17.5%), and c-kithigh population (3.5%) from Ly5.2-expressing mice. The equivalent of 2 × 105 WBM cells from each population was injected into irradiated Ly5.1 recipients in competition with 2 × 105 Ly5.1 WBM cells. Thus, 2 × 105 × 79% or 1.58 × 105 cells of the c-kit− population was injected per recipient, 0.35 × 105 cells of the c-kitlow population per recipient and 0.07 × 105 cells of the c-kithigh population per recipient. Recipient mice were assayed for donor type reconstitution in each lineage after at least 16 weeks' postreconstitution. The percentage of Ly5.2+ T cells, B cells, and myeloid cells was determined by double staining with anti-Ly5.1 and either anti-CD3, B220, Mac-1, or Gr-1. The percentage of donor-derived cells of each lineage are shown for each recipient of c-kit−, c-kitlow, or c-kithigh cells (C). Each line of each column represents the percent of donor-derived cells of each lineage in an individual recipient. WBM from mice that had received a single dose of 5-FU 3 days prior was also collected and treated as described above (D, 5-FU WBM). The distribution of cells in each gate from the lineage− population of 5-FU–treated bone marrow is shown in E (5-FU lin−). The 3 populations were sorted from WBM of 5-FU–treated Ly5.2 mice and 1.95 × 105 cells of the c-kit− cells, 0.05 × 105 of the c-kitlow cells, and 120 of the c-kithigh cells were injected per irradiated Ly5.1 recipient in competition with 2 × 105 Ly5.1 WBM cells. The long-term reconstitution data are shown as the percent of donor-derived cells in each lineage (F ) and represent the sum of 2 independent experiments with 5 mice in each group.

Expression of c-kit on long-term reconstituting stem cells from normal and 5-FU–treated bone marrow. Erythrocyte-depleted WBM from normal mice was stained with anti–c-kit and the lineage cocktail and was sorted into 3 populations as indicated (A, WBM). The relative representation of each population is shown as a percentage within each gate. For comparison, the relative representation of the 3 populations in lineage− bone marrow is shown (B, lin−). WBM cells were sorted based only on the expression of c-kit into a c-kit− population (79% of bone marrow), c-kitlow population (17.5%), and c-kithigh population (3.5%) from Ly5.2-expressing mice. The equivalent of 2 × 105 WBM cells from each population was injected into irradiated Ly5.1 recipients in competition with 2 × 105 Ly5.1 WBM cells. Thus, 2 × 105 × 79% or 1.58 × 105 cells of the c-kit− population was injected per recipient, 0.35 × 105 cells of the c-kitlow population per recipient and 0.07 × 105 cells of the c-kithigh population per recipient. Recipient mice were assayed for donor type reconstitution in each lineage after at least 16 weeks' postreconstitution. The percentage of Ly5.2+ T cells, B cells, and myeloid cells was determined by double staining with anti-Ly5.1 and either anti-CD3, B220, Mac-1, or Gr-1. The percentage of donor-derived cells of each lineage are shown for each recipient of c-kit−, c-kitlow, or c-kithigh cells (C). Each line of each column represents the percent of donor-derived cells of each lineage in an individual recipient. WBM from mice that had received a single dose of 5-FU 3 days prior was also collected and treated as described above (D, 5-FU WBM). The distribution of cells in each gate from the lineage− population of 5-FU–treated bone marrow is shown in E (5-FU lin−). The 3 populations were sorted from WBM of 5-FU–treated Ly5.2 mice and 1.95 × 105 cells of the c-kit− cells, 0.05 × 105 of the c-kitlow cells, and 120 of the c-kithigh cells were injected per irradiated Ly5.1 recipient in competition with 2 × 105 Ly5.1 WBM cells. The long-term reconstitution data are shown as the percent of donor-derived cells in each lineage (F ) and represent the sum of 2 independent experiments with 5 mice in each group.

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