Fig. 4.
Fig. 4. Effects of AdSDF1 on progenitor and stem cells. / (A) Peripheral blood was obtained from each group on day 5 and day 10 after administration of vector. Cells from BM, spleen, and peripheral blood in these mice were seeded in the colony assays and 3 CFU types were scored: CFU-GM (■), BFU-E (░), and CFU-GEMM (▪). (B) Seven days after adenovirus vector administration, 3 mice in each group were humanely killed, and progenitor content was quantified by methylcellulose-based colony assays. (A) and (B) are expressed as the mean (n = 3) ± SEM. Points that differ significantly from data for AdNull administration on the same day are marked: *P < .005, **P < .05. (C) Effects of AdSDF1 on CFU-S in peripheral blood. Peripheral blood was collected from the orbital plexus and pooled before and 1, 3, 5, 7, 10, and 14 days after the onset of treatment. After elimination of erythrocytes, 1 × 105 MNCs from AdSDF1 (▪) or AdNull (░) or normal (■) SCID were injected to irradiated recipient mice. The spleen of the recipients was obtained 12 days later for spleen colony counting. All data are expressed as the mean ± SEM of 3 to 4 experiments. Asterisk (*) indicates results that differ significantly from data for AdNull administration on the same day: *P < .005. (D) Flow cytometry analyses were done in bone marrow MNCs (BMMNCs) 150 days after allogeneic peripheral blood cell transplantation. BMMNCs were stained with H-2Kd–FITC (donor type). The representative percentages of positive populations in BMMNCs are shown. The data from age-matched normal C57BL/6 mice are also represented as a control. (E) Limiting dilution analysis of PBMNCs after administration of adenovirus vector. Lethally irradiated BALB/c mice were transplanted with serial doses of PBMNCs from AdNull- or AdSDF1-treated SCID mice 5 days after vector administration (■, 1 × 106 PBMNCs derived from the peripheral circulation of AdSDF1-treated mice; ▵, 1 × 105 PBMNCs from ADSDF1-treated mice; ♦, 1 × 104 PBMNCs from AdSDF1-treated mice; ○, 1 × 106 PBMNCs from AdNull-treated mice). Survival rate was monitored until 150 days. We used 6 mice in each group. The result from the 1 × 105 injected group compared with AdNull control group achieves statistical significance (**P < .05).

Effects of AdSDF1 on progenitor and stem cells.

(A) Peripheral blood was obtained from each group on day 5 and day 10 after administration of vector. Cells from BM, spleen, and peripheral blood in these mice were seeded in the colony assays and 3 CFU types were scored: CFU-GM (■), BFU-E (░), and CFU-GEMM (▪). (B) Seven days after adenovirus vector administration, 3 mice in each group were humanely killed, and progenitor content was quantified by methylcellulose-based colony assays. (A) and (B) are expressed as the mean (n = 3) ± SEM. Points that differ significantly from data for AdNull administration on the same day are marked: *P < .005, **P < .05. (C) Effects of AdSDF1 on CFU-S in peripheral blood. Peripheral blood was collected from the orbital plexus and pooled before and 1, 3, 5, 7, 10, and 14 days after the onset of treatment. After elimination of erythrocytes, 1 × 105 MNCs from AdSDF1 (▪) or AdNull (░) or normal (■) SCID were injected to irradiated recipient mice. The spleen of the recipients was obtained 12 days later for spleen colony counting. All data are expressed as the mean ± SEM of 3 to 4 experiments. Asterisk (*) indicates results that differ significantly from data for AdNull administration on the same day: *P < .005. (D) Flow cytometry analyses were done in bone marrow MNCs (BMMNCs) 150 days after allogeneic peripheral blood cell transplantation. BMMNCs were stained with H-2Kd–FITC (donor type). The representative percentages of positive populations in BMMNCs are shown. The data from age-matched normal C57BL/6 mice are also represented as a control. (E) Limiting dilution analysis of PBMNCs after administration of adenovirus vector. Lethally irradiated BALB/c mice were transplanted with serial doses of PBMNCs from AdNull- or AdSDF1-treated SCID mice 5 days after vector administration (■, 1 × 106 PBMNCs derived from the peripheral circulation of AdSDF1-treated mice; ▵, 1 × 105 PBMNCs from ADSDF1-treated mice; ♦, 1 × 104 PBMNCs from AdSDF1-treated mice; ○, 1 × 106 PBMNCs from AdNull-treated mice). Survival rate was monitored until 150 days. We used 6 mice in each group. The result from the 1 × 105 injected group compared with AdNull control group achieves statistical significance (**P < .05).

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