Skip to main content
Figure 5 | Journal of Translational Medicine

Figure 5

From: The thrombopoietin receptor, c-Mpl, is a selective surface marker for human hematopoietic stem cells

Figure 5

Human c-mpl+ HSC/PC show significant repopulation of mature single positive CD4+ and CD8+ thymocytes. SCID-hu Thy/Liv grafts (HLA-B8--; n = 21) were sublethally irradiated and injected with 60,000 or 30,000 sorted HLA-disparate (HLA-B8+) CD34+CD38--/dimc-mpl+ or CD34+CD38--/dimc-mpl-- human ABM cells and allowed to repopulate for 8 weeks. Three grafts were reserved as non-injected controls. The surviving Thy/Liv grafts (c-mpl+ = 4; c-mpl-- = 7; Controls = 3) were harvested and single cell suspensions were stained with PI and fluorescence-conjugated antibodies against T-lineage markers, the HLA-B8 donor marker, and appropriate isotype controls. Events (100,000) were acquired and analyzed using FlowJo® software to determine the percentage of viable (PI--) donor-derived (HLA-B8+) MNC expressing the different T-lineage markers from among all viable MNC expressing that particular marker. Viable (PI--) MNC events from the non-injected Thy/Liv grafts were analyzed to establish the HLA-B8-- gate (representing 99.0% of the left-most events on an HLA-B8-APC log histogram plot) and the HLA-B8+ gate (extending from the end of the HLA-B8-- gate to the far right of the log histogram). These gates were then applied without alteration to the HLA-B8-APC log histogram plots of thymocyte subsets from the c-mpl+ and c-mpl---injected Thy/Liv grafts. A. The mean percentage of viable (PI--) CD2-PE+ HLA-B8+ events, out of the total viable CD2-PE+ MNC population is shown for the non-injected grafts (n = 3), the CD34+CD38--c-mpl---injected grafts (n = 7), and the CD34+CD38--c-mpl+-injected grafts (n = 4). Error bars = ± SEM. B. The mean percentage of viable (PI--) HLA-B8+ donor-derived CD4/CD8 subset thymocytes [CD4+CD8--, CD4--CD8+, CD4+CD8+, CD4--CD8--], out of the respective total thymocyte subset population, is shown for the three treatment arms. Error bars = ± SEM.

Back to article page