Figure 3
Figure 3. Long-term reconstitution of lymphoid progenitors and mature lymphocytes in unconditioned X-SCID mice after fetal transplantation of LMPPs and HSCs. (A-G) Fetal X-SCID mice transplanted with 200 LSK CD34+FLT3hi LMPPs () or 200 LSK CD34−FLT3− HSCs () were analyzed for donor-derived lymphoid progenitors and mature lymphocytes in PB, thymus, BM, and spleen 36 weeks after transplantation. (A) Mean (SEM) donor-derived contribution to total nucleated blood cells (top left panel), B cells (B220+C19+; top right panel), and T cells (CD4+ and CD8+; bottom panel) from 5 reconstituted mice. (B) FACS profiles of PB analysis from typical fetal X-SCID recipient transplanted with 200 LMPPs (top FACS plots) and 200 HSCs (bottom FACS plots). For each of the 2 populations, top panels show donor-derived CD45.1 (top left) B220+CD19+ B cells (top right). Bottom left panel shows frequency of TCR β+ T cells among total CD45.1+ cells, and bottom right panel shows distribution of TCR β+ cells into CD4+ and CD8+ blood cells. Gated numbers represent frequencies of total donor cells. (C) Mean (SEM) donor contribution to total B220 cells (B220+), pre B (B220+CD43−IgM−), and immature/mature B cells (B220+CD43−IgM+) in BM (per 2 femurs and 2 tibias) of 5 reconstituted mice. (D) Mean (SEM) donor contribution to DP, SP CD4, and SP CD8 cells in thymus from 5 mice, transplanted 36 weeks earlier. (E) Representative CD4 and CD8 FACS profiles of thymus (gated on CD45.1+ donor cells) in mice transplanted with LMPPs (ii) and HSCs (iii) 36 weeks earlier, showing variable reconstitution of CD4+CD8+ DP thymocytes, and compared with same profiles in WT control (nontransplanted) mouse (i). Percentages for LMPP and HSC transplanted mice are relative to total donor cells. (F,G) TCR V-beta rearrangement of (F) CD4+ and (G) CD8+ donor (CD45.1+) derived cells in spleen. Bar graphs represent mean (± SEM) percentage of cells (of 6 mice analyzed) expressing different V-β subtypes of total donor (F) CD4 and (G) CD8 cells. For all panels, all differences between LMPP and HSC transplanted mice were nonsignificant (P > .05).

Long-term reconstitution of lymphoid progenitors and mature lymphocytes in unconditioned X-SCID mice after fetal transplantation of LMPPs and HSCs. (A-G) Fetal X-SCID mice transplanted with 200 LSK CD34+FLT3hi LMPPs () or 200 LSK CD34FLT3 HSCs () were analyzed for donor-derived lymphoid progenitors and mature lymphocytes in PB, thymus, BM, and spleen 36 weeks after transplantation. (A) Mean (SEM) donor-derived contribution to total nucleated blood cells (top left panel), B cells (B220+C19+; top right panel), and T cells (CD4+ and CD8+; bottom panel) from 5 reconstituted mice. (B) FACS profiles of PB analysis from typical fetal X-SCID recipient transplanted with 200 LMPPs (top FACS plots) and 200 HSCs (bottom FACS plots). For each of the 2 populations, top panels show donor-derived CD45.1 (top left) B220+CD19+ B cells (top right). Bottom left panel shows frequency of TCR β+ T cells among total CD45.1+ cells, and bottom right panel shows distribution of TCR β+ cells into CD4+ and CD8+ blood cells. Gated numbers represent frequencies of total donor cells. (C) Mean (SEM) donor contribution to total B220 cells (B220+), pre B (B220+CD43IgM), and immature/mature B cells (B220+CD43IgM+) in BM (per 2 femurs and 2 tibias) of 5 reconstituted mice. (D) Mean (SEM) donor contribution to DP, SP CD4, and SP CD8 cells in thymus from 5 mice, transplanted 36 weeks earlier. (E) Representative CD4 and CD8 FACS profiles of thymus (gated on CD45.1+ donor cells) in mice transplanted with LMPPs (ii) and HSCs (iii) 36 weeks earlier, showing variable reconstitution of CD4+CD8+ DP thymocytes, and compared with same profiles in WT control (nontransplanted) mouse (i). Percentages for LMPP and HSC transplanted mice are relative to total donor cells. (F,G) TCR V-beta rearrangement of (F) CD4+ and (G) CD8+ donor (CD45.1+) derived cells in spleen. Bar graphs represent mean (± SEM) percentage of cells (of 6 mice analyzed) expressing different V-β subtypes of total donor (F) CD4 and (G) CD8 cells. For all panels, all differences between LMPP and HSC transplanted mice were nonsignificant (P > .05).

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