scholarly journals A wave of WNT signaling balanced by secreted inhibitors controls primitive streak formation in micropattern colonies of human embryonic stem cells

Development ◽  
2019 ◽  
Vol 146 (6) ◽  
pp. dev172791 ◽  
Author(s):  
Iain Martyn ◽  
Ali H. Brivanlou ◽  
Eric D. Siggia
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Wnt Signaling ◽  
Human Embryonic Stem Cells ◽  
Embryonic Stem ◽  
Primitive Streak

scholarly journals Coordination of sonic hedgehog and Wnt signaling determines ventral and dorsal telencephalic neuron types from human embryonic stem cells

Development ◽  
2009 ◽  
Vol 136 (23) ◽  
pp. 4055-4063 ◽  
Author(s):  
X.-J. Li ◽  
X. Zhang ◽  
M. A. Johnson ◽  
Z.-B. Wang ◽  
T. LaVaute ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Wnt Signaling ◽  
Sonic Hedgehog ◽  
Human Embryonic Stem Cells ◽  
Embryonic Stem

scholarly journals Wnt signaling promotes hematoendothelial cell development from human embryonic stem cells

Blood ◽  
2008 ◽  
Vol 111 (1) ◽  
pp. 122-131 ◽  
Author(s):  
Petter S. Woll ◽  
Julie K. Morris ◽  
Matt S. Painschab ◽  
Rebecca K. Marcus ◽  
Aimee D. Kohn ◽  
...  
Keyword(s):  
Stem Cells ◽  
Endothelial Cells ◽  
Embryonic Stem Cells ◽  
Wnt Signaling ◽  
Human Embryonic Stem Cells ◽  
Stromal Cells ◽  
Hematopoietic Cells ◽  
Embryonic Stem ◽  
Canonical Wnt Signaling ◽  
Canonical Wnt

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 CD34brightCD31+Flk1+ cells and a later population of CD34dimCD45+ cells. While the CD34brightCD31+Flk1+ have a more complex morphology and can develop into both endothelial cells and hematopoietic cells, the CD34dimCD45+ 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 CD34brightCD31+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.

Differentiation of human embryonic stem cells into cone photoreceptors through simultaneous inhibition of BMP, TGFβ and Wnt signaling

Development ◽  
2015 ◽  
Vol 142 (19) ◽  
pp. 3294-3306 ◽  
Author(s):  
Shufeng Zhou ◽  
Anthony Flamier ◽  
Mohamed Abdouh ◽  
Nicolas Tétreault ◽  
Andrea Barabino ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Wnt Signaling ◽  
Human Embryonic Stem Cells ◽  
Embryonic Stem ◽  
Cone Photoreceptors ◽  
Simultaneous Inhibition

Targeting a GFP reporter gene to the MIXL1 locus of human embryonic stem cells identifies human primitive streak–like cells and enables isolation of primitive hematopoietic precursors

Blood ◽  
2008 ◽  
Vol 111 (4) ◽  
pp. 1876-1884 ◽  
Author(s):  
Richard P. Davis ◽  
Elizabeth S. Ng ◽  
Magdaline Costa ◽  
Anna K. Mossman ◽  
Koula Sourris ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Human Embryonic Stem Cells ◽  
Fluorescent Protein ◽  
Embryonic Stem ◽  
Growth Factor Receptor ◽  
Primitive Streak ◽  
Factor Receptor Alpha ◽  
Cell Surface Staining ◽  
Green Fluorescent

Differentiating human embryonic stem cells (HESCs) represent an experimental platform for establishing the relationships between the earliest lineages that emerge during human development. Here we report the targeted insertion in HESCs of sequences encoding green fluorescent protein (GFP) into the locus of MIXL1, a gene transiently expressed in the primitive streak during embryogenesis.1,2 GFP fluorescence in MIXL1GFP/w HESCs differentiated in the presence of BMP4 reported the expression of MIXL1, permitting the identification of viable human primitive streak-like cells. The use of GFP as a reporter for MIXL1 combined with cell surface staining for platelet-derived growth factor receptor alpha (PDGFRα) enabled the isolation of a cell population that was highly enriched in primitive hematopoietic precursors, the earliest derivatives of the primitive streak. These experiments demonstrate the utility of MIXL1GFP/w HESCs for analyzing the previously inaccessible events surrounding the development of human primitive streak-like cells and their subsequent commitment to hematopoiesis.

scholarly journals Noncanonical Wnt Signaling Orchestrates Early Developmental Events toward Hematopoietic Cell Fate from Human Embryonic Stem Cells

2009 ◽  
Vol 4 (3) ◽  
pp. 248-262 ◽  
Author(s):  
Kausalia Vijayaragavan ◽  
Eva Szabo ◽  
Marc Bossé ◽  
Veronica Ramos-Mejia ◽  
Randall T. Moon ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Wnt Signaling ◽  
Cell Fate ◽  
Human Embryonic Stem Cells ◽  
Embryonic Stem ◽  
Hematopoietic Cell ◽  
Early Developmental

scholarly journals The negative impact of Wnt signaling on megakaryocyte and primitive erythroid progenitors derived from human embryonic stem cells

2014 ◽  
Vol 12 (2) ◽  
pp. 441-451 ◽  
Author(s):  
Prasuna Paluru ◽  
Kristin M. Hudock ◽  
Xin Cheng ◽  
Jason A. Mills ◽  
Lei Ying ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Wnt Signaling ◽  
Human Embryonic Stem Cells ◽  
Negative Impact ◽  
Embryonic Stem ◽  
Erythroid Progenitors

scholarly journals Generation of qualified clinical-grade functional hepatocytes from human embryonic stem cells in chemically defined conditions

2019 ◽  
Vol 10 (10) ◽  
Author(s):  
Zhongwen Li ◽  
Jun Wu ◽  
Lei Wang ◽  
Weifang Han ◽  
Juan Yu ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Human Embryonic Stem Cells ◽  
Embryonic Stem ◽  
Primitive Streak ◽  
Good Manufacturing Practice ◽  
Glycogen Storage ◽  
Clinical Grade ◽  
Cytochrome P450 Activity

Abstract Hepatocytes have been successfully generated from human pluripotent stem cells (hPSCs). However, the cost-effective and clinical-grade generation of hepatocytes from hPSCs still need to be improved. In this study, we reported the production of functional hepatocytes from clinical-grade human embryonic stem cells (hESCs) under good manufacturing practice (GMP) requirements. We sequentially generated primitive streak (PS), definitive endoderm (DE), hepatoblasts and hepatocyte-like cells (HLCs) from hESCs in the different stages with completely defined reagents. During hepatoblast differentiation, dimethyl sulfoxide (DMSO), transferrin, L-ascorbic acid 2-phosphate sesquimagnesium salt hydrate (Vc-Mg), insulin, and sodium selenite were used instead of cytokines and FBS/KOSR. Then, hepatoblasts were differentiated into HLCs that had a typical hepatocyte morphology and possessed characteristics of mature hepatocytes, such as metabolic-related gene expression, albumin secretion, fat accumulation, glycogen storage, and inducible cytochrome P450 activity in vitro. HLCs integrated into the livers of Tet-uPA Rag2–/– Il2rg–/– (URG) mice, which partially recovered after transplantation. Furthermore, a series of biosafety-related experiments were performed to ensure future clinical applications. In conclusion, we developed a chemically defined system to generate qualified clinical-grade HLCs from hESCs under GMP conditions. HLCs have been proven to be safe and effective for treating liver failure. This efficient platform could facilitate the treatment of liver diseases using hESC-derived HLCs transplantation.

Forced aggregation of defined numbers of human embryonic stem cells into embryoid bodies fosters robust, reproducible hematopoietic differentiation

Blood ◽  
2005 ◽  
Vol 106 (5) ◽  
pp. 1601-1603 ◽  
Author(s):  
Elizabeth S. Ng ◽  
Richard P. Davis ◽  
Lisa Azzola ◽  
Edouard G. Stanley ◽  
Andrew G. Elefanty
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Human Embryonic Stem Cells ◽  
Therapeutic Potential ◽  
Embryonic Stem ◽  
Green Fluorescence Protein ◽  
Primitive Streak ◽  
Embryoid Bodies ◽  
Hematopoietic Growth Factors ◽  
Uniform Size

Abstract To realize the therapeutic potential of human embryonic stem cells (hESCs), it is necessary to regulate their differentiation in a uniform and reproducible manner. We have developed a method in which known numbers of hESCs in serum-free medium were aggregated by centrifugation to foster the formation of embryoid bodies (EBs) of uniform size (spin EBs). These spin EBs differentiated efficiently and synchronously, as evidenced by the sequential expression of molecular markers representing stem cells, primitive streak, and mesoderm. In the presence of hematopoietic growth factors, reproducible differentiation was achieved with blood cells formed in more than 90% of EBs. Using chimeric EBs generated from mixtures of green fluorescence protein–positive (GFP+) and GFP– hESCs in a clonogenic assay, hematopoietic precursor frequency was estimated to be approximately 1:500 input cells. This method of EB formation provides a generally applicable means for modulating and objectively monitoring the directed differentiation of hESCs.

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