Figure 2.
Figure 2. Enhanced donor cell homing and long-term engraftment following fetal HSC mobilization and IUHCT without detrimental effects on fetal recipients. B6GFP BM cells (10 × 106) were injected into E14 Balb/c fetuses via the vitelline vein 60 minutes after maternal administration of vehicle, AMD3100, firategrast, or firategrast+AMD3100. (A) The percentage of donor lineage- (CD3−B220−Gr1−CD11b−) Sca-1+c-Kit+ cells out of total donor CD45+ cells in the FL 24 hours after IUHCT was assessed by flow cytometry. (B) The peripheral blood (PB) of recipients of IUHCT ± mobilizing agent was assessed monthly from 1 to 6 months of life for total donor chimerism defined as the %CD45+ cells that were GFP+. (C) At 6 to 9 months of age, the PB was assessed for multilineage engraftment by determining the percentage of donor cells that express CD3 (T cells), B220 (B cells), CD11b (macrophages/monocytes), and Gr1 (granulocytes). Of note, surface markers, including CD11b and Gr1, are expressed by other cell lineages in addition to macrophages and granulocytes, thus explaining the total percentage of lineage cells >100%. (D) Analysis of the percentage of the CD45+ cells that are GFP+Lineage+ suggests that increased engraftment after treatment with firategrast or firategrast+AMD3100 results from an increase in donor cells of all lineages. (★P < .05 vs vehicle for A-E; see supplemental Figure 5 for lineage flow cytometry plots). (E) Survival to birth was assessed in recipients of IUHCT ± mobilizing agent, and no significant difference was noted between groups (P = .6; values represent the mean survival of 4-5 injected litters per group, n represents total number of fetuses injected per group). (F) Recipients of IUHCT ± mobilizing agent were weighed monthly from 1 to 6 months of life, and no significant difference was noted between treatment groups and the control group (P = .52). At the time of weighing, recipients were also evaluated and demonstrated no evidence of GVHD including ruffled fur, mucositis, fur loss, and hunched posture (data not shown).

Enhanced donor cell homing and long-term engraftment following fetal HSC mobilization and IUHCT without detrimental effects on fetal recipients. B6GFP BM cells (10 × 106) were injected into E14 Balb/c fetuses via the vitelline vein 60 minutes after maternal administration of vehicle, AMD3100, firategrast, or firategrast+AMD3100. (A) The percentage of donor lineage- (CD3−B220−Gr1−CD11b−) Sca-1+c-Kit+ cells out of total donor CD45+ cells in the FL 24 hours after IUHCT was assessed by flow cytometry. (B) The peripheral blood (PB) of recipients of IUHCT ± mobilizing agent was assessed monthly from 1 to 6 months of life for total donor chimerism defined as the %CD45+ cells that were GFP+. (C) At 6 to 9 months of age, the PB was assessed for multilineage engraftment by determining the percentage of donor cells that express CD3 (T cells), B220 (B cells), CD11b (macrophages/monocytes), and Gr1 (granulocytes). Of note, surface markers, including CD11b and Gr1, are expressed by other cell lineages in addition to macrophages and granulocytes, thus explaining the total percentage of lineage cells >100%. (D) Analysis of the percentage of the CD45+ cells that are GFP+Lineage+ suggests that increased engraftment after treatment with firategrast or firategrast+AMD3100 results from an increase in donor cells of all lineages. (★P < .05 vs vehicle for A-E; see supplemental Figure 5 for lineage flow cytometry plots). (E) Survival to birth was assessed in recipients of IUHCT ± mobilizing agent, and no significant difference was noted between groups (P = .6; values represent the mean survival of 4-5 injected litters per group, n represents total number of fetuses injected per group). (F) Recipients of IUHCT ± mobilizing agent were weighed monthly from 1 to 6 months of life, and no significant difference was noted between treatment groups and the control group (P = .52). At the time of weighing, recipients were also evaluated and demonstrated no evidence of GVHD including ruffled fur, mucositis, fur loss, and hunched posture (data not shown).

Close Modal
This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal