Figure 2. PPARγ antagonism expands long term HSCs from human cord blood.

(a) Representative FACS plots showing the percent engraftment of vehicle control- or GW9662-treated human CB CD34⁺ cells in the bone marrow of recipient NSG mice 4 months after transplantation. Non-transplanted mouse bone marrow cells were used as the negative control. Representative plots from 3 independent experiments are shown.

(b) The percentage of human CD45⁺ cells in peripheral blood (PB) at 2 and 4 months, and the percentage of human CD45⁺ cells in bone marrow (BM) at 4 months, after transplantation of NSG mice with the progeny of 30,000 CB CD34⁺ cells treated with vehicle or GW9662 for 4 days (n=5 mice per group). Data are shown as dot plots (mean±s.e.m.). *p<0.05, **p=0.009.

(c–e) Human CD33⁺ myeloid cell, CD3⁺ T cell and CD19⁺ B cell chimerism in the bone marrow (BM) of primary recipients at 4 months after transplantation (n=5 mice per group). Data are shown as dot plots (mean±s.e.m.). *p=0.032, **p<0.01.

(f) The engraftment of human CD45⁺ cells in the peripheral blood and bone marrow of secondary recipient NSG mice at 4 months after transplantation (n=5 mice per group). Data are shown as dot plots (mean±s.e.m.). From (b) to (f), two-tailed Student’s t-test was used. *p=0.017, **p=0.002.

(g,h) The frequency of human SRCs in uncultured CB CD34⁺ cells or in the progeny of an equivalent number of CD34⁺ cells treated by vehicle or GW9662 for 4 days. Graded doses of uncultured, vehicle or GW9662 treated CB CD34⁺ cells were transplanted into irradiated NSG mice and the percentage of human CD45⁺ cells in bone marrow (BM) was analyzed at 3 months after transplantation (n=2–5 mice per group, see Supplementary Table 2). Poisson statistical analysis of data from Supplementary Table 2 and 3 is shown in (g). The percentage of negative mice for each dose of cells is plotted. Inverted triangles indicate that all tested mice were positive in this group. Solid lines indicate the best-fit linear model for each data set. Group A (black line) corresponds to the uncultured group, group B (red line) corresponds to the vehicle-treated group, and group C (green line) corresponds to the GW9662-treated group. Dotted lines represent 95% confidence intervals. HSC frequencies (line in the box) and 95% confidence intervals (box) presented as the number of SRCs in 1 × 10⁶ CD34⁺ cells are shown in (h). NS, not significant; *p=0.0375, **p=0.00828. Results from another independent experiment are shown in Supplementary Figure 6b,c and Supplementary Tables 4 and 5.