Germ Layer Induction in ESC—Following the Vertebrate Roadmap

Smith, James C.; Wardle, Fiona C.; Loose, Matthew; Stanley, Edouard G.; Patient, Roger

doi:10.1002/9780470151808.sc01d01s1

Cited by 5 publications

(2 citation statements)

References 212 publications

(280 reference statements)

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“…Following the expression of these genes, induction of markers representing differentiated cell types can be observed, such as Pdx1 (foregut endoderm), Nkx2-5 (cardiac mesoderm) and β H1 globin (yolk sac erythroid cells) [2] . Thus, parallels exist between the differentiation pathways used by ESCs in vitro and the developmental roadmap followed by cells during the early stages of embryogenesis [4] .…”

Section: Introductionmentioning

confidence: 99%

Differentiating Embryonic Stem Cells Pass through ‘Temporal Windows’ That Mark Responsiveness to Exogenous and Paracrine Mesendoderm Inducing Signals

2010

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BackgroundMesendoderm induction during embryonic stem cell (ESC) differentiation in vitro is stimulated by the Transforming Growth Factor and Wingless (Wnt) families of growth factors.Principal FindingsWe identified the periods during which Bone Morphogenetic Protein (BMP) 4, Wnt3a or Activin A were able to induce expression of the mesendoderm marker, Mixl1, in differentiating mouse ESCs. BMP4 and Wnt3a were required between differentiation day (d) 1.5 and 3 to most effectively induce Mixl1, whilst Activin A induced Mixl1 expression in ESC when added between d2 and d4, indicating a subtle difference in the requirement for Activin receptor signalling in this process. Stimulation of ESCs with these factors at earlier or later times resulted in little Mixl1 induction, suggesting that the differentiating ESCs passed through ‘temporal windows’ in which they sequentially gained and lost competence to respond to each growth factor. Inhibition of either Activin or Wnt signalling blocked Mixl1 induction by any of the three mesendoderm-inducing factors. Mixing experiments in which chimeric EBs were formed between growth factor-treated and untreated ESCs revealed that BMP, Activin and Wnt signalling all contributed to the propagation of paracrine mesendoderm inducing signals between adjacent cells. Finally, we demonstrated that the differentiating cells passed through ‘exit gates’ after which point they were no longer dependent on signalling from inducing molecules for Mixl1 expression.ConclusionsThese studies suggest that differentiating ESCs are directed by an interconnected network of growth factors similar to those present in early embryos and that the timing of growth factor activity is critical for mesendoderm induction.

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Section: Introductionmentioning

confidence: 99%

Differentiating Embryonic Stem Cells Pass through ‘Temporal Windows’ That Mark Responsiveness to Exogenous and Paracrine Mesendoderm Inducing Signals

2010

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“…They sequentially express cohorts of genes that mark specific stages of postimplantation embryonic development, starting from the inner cell mass stage and passing through a series of developmental fate restrictions before expressing genes that mark derivatives of mesoderm, endoderm, and ectoderm germ layers (Keller et al, 1993;Robertson et al, 2000;Lacaud et al, 2002;Ng et al, 2005a,b;Hirst et al, 2006). This developmental concordance between embryonic development and in vitro ESC differentiation is reflected by the fact that the same growth factors that induce and pattern the embryo also allow directed differentiation of ESC (Smith et al, 2007;UNIT 1D.1).…”

Section: Background Informationmentioning

confidence: 99%

Directed Differentiation of Human Embryonic Stem Cells as Spin Embryoid Bodies and a Description of the Hematopoietic Blast Colony Forming Assay

Davis

Hatzistavrou

et al. 2008

CP Stem Cell Biology

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This unit describes a protocol for the differentiation of human embryonic stem cells (hESCs). To generate spin embryoid bodies (EBs), known numbers of hESCs are deposited into low‐attachment, round‐bottomed 96‐well plates in a serum‐free medium supplemented with growth factors. The cells are then aggregated by centrifugation, initiating formation of EBs of uniform size. The spin EBs generated using this technique differentiate efficiently and synchronously along the lineages preferentially induced by the combinations of growth factors to which the cells are exposed. The 96‐well format permits an assessment of the effects of different combinations of growth factors in the same experiment, facilitating the optimization of differentiation conditions for any given cell type. Up to 40 plates can be set up within a couple of hours by one experimenter, and aliquots of the differentiating EBs can be harvested at intervals and subjected to analyses using a variety of techniques. Curr. Protoc. Stem Cell Biol. 4:1D.3.1‐1D.3.23. © 2008 by John Wiley & Sons, Inc.

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Stem cell decision making and critical-like exploratory networks

2009

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A sound theoretical or conceptual model of gene regulatory processes that control stem cell fate is still lacking, compromising our ability to manipulate stem cells for therapeutic benefit. The complexity of the regulatory and signaling pathways limits development of useful, predictive models that employ solely reductionist methods using molecular components. However, there is clear evidence from other complex systems that coarse-grained or mesoscale models can yield useful insights and provide workable models for the prediction of some emergent properties such as cell phenotype. We present such a coarse-grained model of stem cell decision making, utilizing the concept of self-organized criticality, which is an order that propagates in some nonequilibrium systems. The model proposes that stochastic gene expression within a stem cell gene regulatory network self-organizes to a critical-like state, characterized by cascades of gene expression that prime various transcriptional programs associated with different cell fates. This diversity of cell fate options is reduced during the decision-making process, which involves a supercritical connectivity in the gene regulatory network as a stem cell leaves its niche microenvironment and an overall increase in transcription occurs. As modules of genes that correspond to specific cell fates approach their critical points, competitive interactions occur between them that are influenced by prevailing microenvironmental conditions. The conceptual model incorporates both intrinsic and extrinsic factors governing stem cell fate and provides a logical pathway to the development of a computational model. We further suggest that rapid self-organized criticality, rather than self-organized criticality, best describes the mesoscale organization of gene regulatory networks.

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Germ Layer Induction in ESC—Following the Vertebrate Roadmap

Cited by 5 publications

References 212 publications

Differentiating Embryonic Stem Cells Pass through ‘Temporal Windows’ That Mark Responsiveness to Exogenous and Paracrine Mesendoderm Inducing Signals

Differentiating Embryonic Stem Cells Pass through ‘Temporal Windows’ That Mark Responsiveness to Exogenous and Paracrine Mesendoderm Inducing Signals

Directed Differentiation of Human Embryonic Stem Cells as Spin Embryoid Bodies and a Description of the Hematopoietic Blast Colony Forming Assay

Stem cell decision making and critical-like exploratory networks

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