American Society of Hematology

Figure 4.

$Figure 4. In vivo assays of cells maintained in vitro with NCCs or sorted fractions of NCCs. (A) Schema summarizing the protocol for maintaining LSK cells in cocultures followed by transplantation of progeny cells into CD45.1/CD45.2 recipients. (B) Progeny of 1000 LSK cells from C57BL/6 (CD45.2) mice cocultured for 5 days with each group of cells identified in panel A were transplanted in a competitive repopulation assay with 100 000 BoyJ (CD45.1) cells via tail vein injection in lethally irradiated (split dose of 650 and 350 cGy) CD45.1 × CD45.2 F1 recipients. Freshly isolated LSK cells from BoyJ mice were used as control. Engraftment was assessed in the peripheral blood monthly and plotted for each group (n = 8–9 mice per group from 2 independent experiments). *P < .05 vs 1w NCC, 1-way ANOVA. (C) Chimerism in the BM of primary recipients (shown in panel B) at 4 months posttransplantation. (D) At 4 months post–primary transplantation, halves of femurs from primary recipients were transplanted into lethally irradiated secondary recipients. Chimerism in secondary recipients at 3 months posttransplantation is shown (4 mice per group) *P < .05 vs LSK group, +P < .05 vs 1w NCC+MK group, 1-way ANOVA.$

In vivo assays of cells maintained in vitro with NCCs or sorted fractions of NCCs. (A) Schema summarizing the protocol for maintaining LSK cells in cocultures followed by transplantation of progeny cells into CD45.1/CD45.2 recipients. (B) Progeny of 1000 LSK cells from C57BL/6 (CD45.2) mice cocultured for 5 days with each group of cells identified in panel A were transplanted in a competitive repopulation assay with 100 000 BoyJ (CD45.1) cells via tail vein injection in lethally irradiated (split dose of 650 and 350 cGy) CD45.1 × CD45.2 F1 recipients. Freshly isolated LSK cells from BoyJ mice were used as control. Engraftment was assessed in the peripheral blood monthly and plotted for each group (n = 8–9 mice per group from 2 independent experiments). *P < .05 vs 1w NCC, 1-way ANOVA. (C) Chimerism in the BM of primary recipients (shown in panel B) at 4 months posttransplantation. (D) At 4 months post–primary transplantation, halves of femurs from primary recipients were transplanted into lethally irradiated secondary recipients. Chimerism in secondary recipients at 3 months posttransplantation is shown (4 mice per group) *P < .05 vs LSK group, ⁺P < .05 vs 1w NCC+MK group, 1-way ANOVA.

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