Figure 6.
A DPT-KO mouse displays improved homing and long-term engraftment. (A) Homing of donor cells in DPT-KO mice was assessed 16 hours after transplant of 2 million CD45.1 donor WBM cells (n = 12-14 animals per group). (B) Homing in DPT-KO animals, with or without 2 µg/mL rDPT delivery 30 minutes prior to adoptive transfer. Homing after HCT was assessed as in (A). (C) Spleen CFU assay. Two million donor cells were adoptively transferred to irradiated recipients. Eight days after the transplant, spleens were harvested and fixed in Bouin’s solution (middle and right panels), and CFU were enumerated (left panel). Scale bar, 5 mm. (D) Nonradiation transplant paradigm for (R-S). Enumeration of homed donor CD45+ marrow cells (E) and LSK marrow cells (F) in 2 leg bones measured 16 hours after transplant. (G) Sublethal transplant paradigm for (E-H). (H) Peripheral blood (PB) donor engraftment measured 20 weeks after transplant (n = 9 or 10 animals per group). (I) Donor bone marrow (BM) engraftment measured 20 weeks after transplant. (J) Donor LSK cells measured 20 weeks after transplant. (K) Donor LSK-SLAM HSPCs measured 20 weeks after transplant. The flow cytometry gating strategy for (G-H) is shown in supplemental Figure 7A. (L) Sublethal transplant paradigm for panels (N-P). LSK HSPCs were isolated by fluorescence-activated cell sorting (FACS) prior to adoptive transfer into irradiated recipients. (M) Peripheral blood mononuclear cell (PBMC) donor engraftment measured 20 weeks after transplant. (N) Percentage of donor marrow LSK cells in transplanted animals. Four leg bones were harvested for assessment 20 weeks after HCT. (O) Secondary HCT was performed with the primary HCT recipients as donors. Peripheral blood donor engraftment was measured 20 weeks after the secondary transplant. (P) Lethal transplant paradigm for (J-L). LSK HSPCs were isolated by FACS prior to mixture with rescue cells and adoptive transfer into irradiated recipients. (Q) Peripheral blood donor engraftment measured 20 weeks after transplant. (R) Percentage of donor marrow LSK cells in transplanted animals. Four leg bones were harvested for assessment 20 weeks after HCT. (S) Percentage of donor LSK-SLAM HSPCs in transplanted animals. Four leg bones were harvested for assessment 20 weeks after HCT. All data are means and SD. The P values were derived using a Student's t test.

A DPT-KO mouse displays improved homing and long-term engraftment. (A) Homing of donor cells in DPT-KO mice was assessed 16 hours after transplant of 2 million CD45.1 donor WBM cells (n = 12-14 animals per group). (B) Homing in DPT-KO animals, with or without 2 µg/mL rDPT delivery 30 minutes prior to adoptive transfer. Homing after HCT was assessed as in (A). (C) Spleen CFU assay. Two million donor cells were adoptively transferred to irradiated recipients. Eight days after the transplant, spleens were harvested and fixed in Bouin’s solution (middle and right panels), and CFU were enumerated (left panel). Scale bar, 5 mm. (D) Nonradiation transplant paradigm for (R-S). Enumeration of homed donor CD45+ marrow cells (E) and LSK marrow cells (F) in 2 leg bones measured 16 hours after transplant. (G) Sublethal transplant paradigm for (E-H). (H) Peripheral blood (PB) donor engraftment measured 20 weeks after transplant (n = 9 or 10 animals per group). (I) Donor bone marrow (BM) engraftment measured 20 weeks after transplant. (J) Donor LSK cells measured 20 weeks after transplant. (K) Donor LSK-SLAM HSPCs measured 20 weeks after transplant. The flow cytometry gating strategy for (G-H) is shown in supplemental Figure 7A. (L) Sublethal transplant paradigm for panels (N-P). LSK HSPCs were isolated by fluorescence-activated cell sorting (FACS) prior to adoptive transfer into irradiated recipients. (M) Peripheral blood mononuclear cell (PBMC) donor engraftment measured 20 weeks after transplant. (N) Percentage of donor marrow LSK cells in transplanted animals. Four leg bones were harvested for assessment 20 weeks after HCT. (O) Secondary HCT was performed with the primary HCT recipients as donors. Peripheral blood donor engraftment was measured 20 weeks after the secondary transplant. (P) Lethal transplant paradigm for (J-L). LSK HSPCs were isolated by FACS prior to mixture with rescue cells and adoptive transfer into irradiated recipients. (Q) Peripheral blood donor engraftment measured 20 weeks after transplant. (R) Percentage of donor marrow LSK cells in transplanted animals. Four leg bones were harvested for assessment 20 weeks after HCT. (S) Percentage of donor LSK-SLAM HSPCs in transplanted animals. Four leg bones were harvested for assessment 20 weeks after HCT. All data are means and SD. The P values were derived using a Student's t test.

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