Figure 5
In vivo depletion of macrophages with clodronate-loaded liposomes mobilizes HSCs. (A) Effect of a 2-day treatment with clo-lip and PBS-lip on the number of BM F4/80+Ly-6G−CD11b+ (F+G−), F4/80+Ly-6G+CD11b+ (F+G+) macrophages and F4/80−Ly-6G+CD11b+ (F−G+) granulocytes measured by flow cytometry. The 2 dot-plots illustrate the loss of F+G+. Data are mean ± SD of 4 mice per group. (B) Effect of liposomes on the number of circulating Lin−Sca1+KIT+ HSPCs and CFCs. Data are mean ± SD of 4 mice per group. (C) Mobilization of long-term reconstituting HSCs in competitive repopulation assay. Test donor CD45.2+ mice were injected with clo-lip for 2 or 4 days, or PBS-lip for 4 days. Next, 50 μL blood was transplanted in competition with 200 000 competing CD45.1+ BM cells into lethally irradiated CD45.1+ recipients. Donor CD45.2+ contribution was measured in CD45+ leukocytes, myeloid, B, and T cells 16 weeks after transplantation. Contribution was estimated to be positive when the proportion of CD45.2+ donor cells was above 0.5% in each lineage. ***P < .001, **P < .01, and *P < .05 between PSB-lip– and clo-lip–treated mice.

In vivo depletion of macrophages with clodronate-loaded liposomes mobilizes HSCs. (A) Effect of a 2-day treatment with clo-lip and PBS-lip on the number of BM F4/80+Ly-6GCD11b+ (F+G), F4/80+Ly-6G+CD11b+ (F+G+) macrophages and F4/80Ly-6G+CD11b+ (FG+) granulocytes measured by flow cytometry. The 2 dot-plots illustrate the loss of F+G+. Data are mean ± SD of 4 mice per group. (B) Effect of liposomes on the number of circulating LinSca1+KIT+ HSPCs and CFCs. Data are mean ± SD of 4 mice per group. (C) Mobilization of long-term reconstituting HSCs in competitive repopulation assay. Test donor CD45.2+ mice were injected with clo-lip for 2 or 4 days, or PBS-lip for 4 days. Next, 50 μL blood was transplanted in competition with 200 000 competing CD45.1+ BM cells into lethally irradiated CD45.1+ recipients. Donor CD45.2+ contribution was measured in CD45+ leukocytes, myeloid, B, and T cells 16 weeks after transplantation. Contribution was estimated to be positive when the proportion of CD45.2+ donor cells was above 0.5% in each lineage. ***P < .001, **P < .01, and *P < .05 between PSB-lip– and clo-lip–treated mice.

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