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. Author manuscript; available in PMC: 2019 Dec 1.
Published in final edited form as: Exp Hematol. 2018 Oct 31;68:2–9. doi: 10.1016/j.exphem.2018.10.009

Figure 1. Isolation and functional characterization of hemogenic endothelial (HE) and endothelial (E) cells from the arteries and yolk sac.

Figure 1.

A) FACS gating strategy to purify HE and E cells from the arteries. Shown are profiles from E10.5 embryos.

B) Purification of HE and E from yolk sacs (YS), E10.5.

C) Frequency of cells that produced endothelial tubes on OP9 cells cultured with vascular endothelial growth factor (VEGF). Data from E10.5 HE and E are shown. Average frequencies are indicated above the bars. Significance determined by ANOVA and Tukey’s multiple comparisons test (mean ± 95% CI, *P < 0.05, **P < 0.01, n=3 experiments).

D) Frequency of HE and E cells that produced CD45+ hematopoietic cells following 8–10 days of culture on OP9 stromal cells in the presence of stem cell factor (SCF), interleukin 3 (IL-3), FLT3 ligand (Flt3L) and IL-7. Data from E10.5 HE and E are shown. Significance determined by ANOVA and Tukey’s multiple comparison tests (mean ± 95% CI, **P < 0.01, n=3 experiments).

E) Methylcellulose assays to enumerate colony forming units - culture (CFU-C) in sorted populations of endothelial cells, and unfractionated yolk sac cells as a control, represented as the number of CFU-Cs per embryo equivalent. Data from E10.5 samples shown. Significance determined by ANOVA and Tukey’s multiple comparison tests (mean ± SD, P < 0.0001, n = 4–8 samples, data combined from 3 experiments).