Germ layer induction from embryonic stem cells

Gadue, Paul; Huber, Tara L.; Nostro, M. Cristina; Kattman, Steven J.; Keller, Gordon

doi:10.1016/j.exphem.2005.06.009

Cited by 117 publications

(108 citation statements)

References 110 publications

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“…In mouse ESCs, both AF have been described as key players in the development of the hematopoietic mesoderm [54][55][56][57][58]. However, aside from BMP4, whose contribution to the formation of hematopoietic mesoderm has been demonstrated [37][38][39], the role of Activin A in early hESC fate decisions has been considerably less explored, apart from The linear regression analysis of the previous 2 parameters post-transfection with Brachyury (F) and scramble (G) siRNAs.…”

Section: Discussionmentioning

confidence: 99%

Activin A Promotes Hematopoietic Fated Mesoderm Development Through Upregulation of Brachyury in Human Embryonic Stem Cells

Cerdan

McIntyre

Mechael

et al. 2012

Stem Cells and Development

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The development of the hematopoietic system involves multiple cellular steps beginning with the formation of the mesoderm from the primitive streak, followed by emergence of precursor populations that become committed to either the endothelial or hematopoietic lineages. A number of growth factors such as activins and fibroblast growth factors (FGFs) are known to regulate the early specification of hematopoietic fated mesoderm, notably in amphibians. However, the potential roles of these factors in the development of mesoderm and subsequent hematopoiesis in the human have yet to be delineated. Defining the cellular and molecular mechanisms by which combinations of mesoderm-inducing factors regulate this stepwise process in human cells in vitro is central to effectively directing human embryonic stem cell (hESC) hematopoietic differentiation. Herein, using hESCderived embryoid bodies (EBs), we show that Activin A, but not basic FGF/FGF2 (bFGF), promotes hematopoietic fated mesodermal specification from pluripotent human cells. The effect of Activin A treatment relies on the presence of bone morphogenetic protein 4 (BMP4) and both of the hematopoietic cytokines stem cell factor and fms-like tyrosine kinase receptor-3 ligand, and is the consequence of 2 separate mechanisms occurring at 2 different stages of human EB development from mesoderm to blood. While Activin A promotes the induction of mesoderm, as indicated by the upregulation of Brachyury expression, which represents the mesodermal precursor required for hematopoietic development, it also contributes to the expansion of cells already committed to a hematopoietic fate. As hematopoietic development requires the transition through a Brachyury + intermediate, we demonstrate that hematopoiesis in hESCs is impaired by the downregulation of Brachyury, but is unaffected by its overexpression. These results demonstrate, for the first time, the functional significance of Brachyury in the developmental program of hematopoietic differentiation from hESCs and provide an in-depth understanding of the molecular cues that orchestrate stepwise development of hematopoiesis in a human system.

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Section: Discussionmentioning

confidence: 99%

Activin A Promotes Hematopoietic Fated Mesoderm Development Through Upregulation of Brachyury in Human Embryonic Stem Cells

Cerdan

McIntyre

Mechael

et al. 2012

Stem Cells and Development

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“…Upon exposure to a differentiation signal these cells are capable of multi-lineage differentiation into cells of the three germ layers: ectoderm, mesoderm, and endoderm (2). The term "commitment" is frequently used with regard to achieving a new differentiated phenotype, assayed by changes in morphology and patterns of gene expression (3)(4)(5).…”

mentioning

confidence: 99%

The Role of Histone Acetylation in Regulating Early Gene Expression Patterns during Early Embryonic Stem Cell Differentiation

McCool

Singer

et al. 2007

Journal of Biological Chemistry

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We have examined the role of histone acetylation in the very earliest steps of differentiation of mouse embryonic stem cells in response to withdrawal of leukemia inhibitory factor (LIF) as a differentiation signal. The cells undergo dramatic changes in morphology and an ordered program of gene expression changes representing differentiation to all three germ layers over the first 3-5 days of LIF withdrawal. We observed a global increase in acetylation on histone H4 and to a lesser extent on histone H3 over this time period. Treatment of the cells with trichostatin A (TSA), a histone deacetylase inhibitor, induced changes in morphology, gene expression, and histone acetylation that mimicked differentiation induced by withdrawal of LIF. We examined localized histone acetylation in the regulatory regions of genes that were transcriptionally either active in undifferentiated cells, induced during differentiation, or inactive under all treatments. There was striking concordance in the histone acetylation patterns of specific genes induced by both TSA and LIF withdrawal. Increased histone acetylation in local regions correlated best with induction of gene expression. Finally, TSA treatment did not support the maintenance or progression of differentiation. Upon removal of TSA, the cells reverted to the undifferentiated phenotype. We concluded that increased histone acetylation at specific genes played a role in their expression, but additional events are required for maintenance of differentiated gene expression and loss of the pluripotent state.

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“…24 Therefore, role of upregulated cell-secreted BMP4 on mESC pluripotency 11 and differentiation (in this study) in the MB correlated well with the respective role of exogenous BMP4 in the conventional macro-scale cultures.…”

Section: Inhibition Of Bmp4 Signaling In the Differentiation Cultumentioning

confidence: 66%

“…We selected these soluble factors on the basis of their influence on mESC differentiation: autocrine FGF4 induces neuronal differentiation; exogenous BMP4 inhibits neuronal but induces mesoand endo-dermal differentiation; and Nodal or Activin induces meso-and endo-dermal differentiation. 24 Among these, only BMP4 expression was remarkably upregulated in the MB as compared to the 6WP culture (Fig. 5(e)).…”

Section: Fig 2 Differentiated Cells In the 6wpmentioning

confidence: 99%

“…5(e)), accumulation of other factors in the small volume might have some influence. For example, besides BMP4, accumulation of Nodal and Activin (can induce meso-and endo-dermal differentiation in mESC) 24 in the small volume of the MB might have role in augmenting meso-and endo-dermal differentiation in the MB. However, inhibition of neuronal differentiation together with augmentation of meso-and endo-dermal differentiation and upregulation of BMP4 strongly suggested prominent role of BMP4 in the MB.…”

Section: Inhibition Of Bmp4 Signaling In the Differentiation Cultumentioning

confidence: 99%

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Induction of alternative fate other than default neuronal fate of embryonic stem cells in a membrane-based two-chambered microbioreactor by cell-secreted BMP4

et al. 2012

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Cell-secreted soluble factor signaling in a diffusion dominant microenvironment plays an important role on early stage differentiation of pluripotent stem cells in vivo. In this study, we utilized a membrane-based two-chambered microbioreactor (MB) to differentiate mouse embryonic stem cells (mESCs) in a diffusion dominant microenvironment of the top chamber while providing enough nutrient through the bottom chamber. Speculating that accumulated FGF4 in the small top chamber will augment neuronal differentiation in the MB culture, we first differentiated mESCs for 8 days by using a chemically optimized culture medium for neuronal induction. However, comparison of cellular morphology and expression of neuronal markers in the MB with that in the 6-well plate (6WP) indicated relatively lower neuronal differentiation in the MB culture. Therefore, to investigate whether microenvironment in the MB facilitates non-neuronal differentiation, we differentiated mESCs for 8 days by using chemically defined basal medium. In this case, differentiated cell morphology differed markedly between the MB and 6WP cultures: epithelial sheet-like morphology in the MB, whereas rosette morphology in the 6WP. Expression of markers from the three germ layers indicated lower neuronal but higher meso-and endo-dermal differentiation of mESCs in the MB than the 6WP culture. Moreover, among various cell-secreted soluble factors, BMP4 expression was remarkably upregulated in the MB culture. Inhibition of BMP4 signaling demonstrated that enhanced effect of upregulated BMP4 was responsible for the prominent meso-and endo-dermal differentiation in the MB. However, in the 6WP, downregulated BMP4 had a minimal influence on the differentiation behavior. Our study demonstrated utilization of a microbioreactor to modulate the effect of cell-secreted soluble factors by autoregulation and thereby inducing alternative self-capability of mESCs. Understanding and implementation of autoregulation of soluble factors similar to this study will lead to the development of robust culture systems to control ESC behavior. V C 2012 American Institute of Physics. [http://dx

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Germ layer induction from embryonic stem cells

Cited by 117 publications

References 110 publications

Activin A Promotes Hematopoietic Fated Mesoderm Development Through Upregulation of Brachyury in Human Embryonic Stem Cells

Activin A Promotes Hematopoietic Fated Mesoderm Development Through Upregulation of Brachyury in Human Embryonic Stem Cells

The Role of Histone Acetylation in Regulating Early Gene Expression Patterns during Early Embryonic Stem Cell Differentiation

Induction of alternative fate other than default neuronal fate of embryonic stem cells in a membrane-based two-chambered microbioreactor by cell-secreted BMP4

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