Wnt signaling promotes hematoendothelial cell development from human embryonic stem cells. Blood

. Stem Cell Institute and Department of Medicine, University of Minnesota, Translational Research Facility, 2001 6th St SE, Minneapolis, MN 55455, USA.
Blood (Impact Factor: 10.45). 02/2008; 111(1):122-31. DOI: 10.1182/blood-2007-04-084186
Source: PubMed


Human embryonic stem cells (hESCs) provide an important means to effectively study soluble and cell-bound mediators that regulate development of early blood and endothelial cells in a human model system. Here, several complementary methods are used to demonstrate canonical Wnt signaling is important for development of hESC-derived cells with both hematopoietic and endothelial potential. Analyses using both standard flow cy-tometry, as well the more detailed high-throughput image scanning flow cytometry, characterizes sequential development of distinct early developing CD34(bright)CD31(+)Flk1(+) cells and a later population of CD34(dim)CD45(+) cells. While the CD34(bright)CD31(+)Flk1(+) have a more complex morphology and can develop into both endothelial cells and hematopoietic cells, the CD34(dim)CD45(+) cells have a simpler morphology and give rise to only hematopoietic cells. Treatment with dickkopf1 to inhibit Wnt signaling results in a dramatic decrease in development of cells with hematoendothelial potential. In addition, activation of the canonical Wnt signaling pathway in hESCs by coculture with stromal cells that express Wnt1, but not use of noncanonical Wnt5-expressing stromal cells, results in an accelerated differentiation and higher percentage of CD34(bright)CD31(+)Flk1(+) cells at earlier stages of differentiation. These studies effectively demonstrate the importance of canonical Wnt signaling to mediate development of early hematoendothelial progenitors during human development.

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    • "We tested a wide range of culture conditions involving stimulation with major determinants of endothelial fate, including varying the timing and concentrations of VEGF, BMP4, CHIR-99021 and FGF, based on previous studies describing endothelial differentiation from PSCs (Kennedy et al., 2007, 2012; Choi et al., 2012; Rafii et al., 2013; White et al., 2013) (supplementary material Fig. S7; data not shown). Studies from our lab and others have established a role for Wnt/β-catenin signaling in specifying endothelial commitment (Woll et al., 2008; Palpant et al., 2013; Sturgeon et al., 2014). We observed, however, that addition of the small molecule Wnt/β-catenin agonist CHIR-99021 between days 2 and 5 inhibited endothelial fate specification, indicating that other factors involved in the specification of endothelium tightly orchestrate the dosage of Wnt/β-catenin signaling (supplementary material Fig. S7). "
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