Figure 7.

Endothelins Promote Hematopoiesis in the Mouse and Human Model Systems

(A) CD31, cKit, and EDN1 immunostaining in the Ao of an E10.5 mouse embryo. Arrowheads indicate cKit+EDN1+ IAHCs or individual emerging Hem cells. The images in (A’’) show a magnification of the boxed region in (A’). Scale bars: 50 μm.

(B) Experimental strategies for mouse and human model systems. MC, methocult.

(C) Production of CFU-Cs in E9.5 caudal tissues cultured with each of the test factors (REN1, EDN1, and EDN2) at 10 ng/μL and 100 ng/μL. n = 4 independent experiments.

(D) Proportion of CFU-Mac and CFU-GEMM from (B), normalized to 1; n = 4 independent experiments.

(E) Repopulation with E9.5 caudal part cells cultured with the test factors (REN1, EDN1, and EDN2) and 18 weeks following transplantation (^∗p < 0.05, paired t test, n = 3 independent experiments). The shape on the graph indicates the experiment. The red line shows the upper limit of control repopulation levels). The numbers of long-term repopulating mice (18 weeks) higher than 1% are as follows: CTRL, 0/11; REN1_10, 3/11; REN1_100, 5/12; EDN1_10, 3/11; EDN1_100, 3/12; EDN2_10, 2/11; EDN2_100, 6/12.

(F) Effect of test factors on production of CFU-Cs by human ES cells at 12 days of differentiation. The number of day 14 CFU-Cs per 5,000 plated cells is shown; factors were added at day 8 of culture (^∗∗p < 0.01, paired t test; error bars, ± SEM, n = 4).

(G) Production of CFU-GEMM from (E) (^∗∗p < 0.01, paired t test; error bars, ± SEM).