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Protein Kinase C Mediated Extraembryonic Endoderm Differentiation of Human Embryonic Stem Cells

ABSTRACT: Unlike mouse embryonic stem (ES) cells, which are closely related to the inner cell mass, human ES cells appear to be more closely related to the later primitive ectoderm. For example, human ES cells and primitive ectoderm share a common epithelial morphology, growth factor requirements, and the potential to differentiate to all three embryonic germ layers. However, it has previously been shown that human ES cells can also differentiate to cells expressing markers of trophoblast, an extraembryonic lineage formed before the formation of primitive ectoderm. Here we show that phorbol ester 12-O-Tetradecanoylphorbol 13-acetate (TPA) causes human ES cells to undergo an epithelial mesenchymal transition and to differentiate into cells expressing markers of parietal endoderm, another extraembryonic lineage. We further confirmed that this differentiation is through the activation of PKC pathway and demonstrated that a particular PKC subtype, PKC-delta, is most responsible for this transition.

ORGANISM(S): Homo sapiens

PROVIDER: GSE33180 | GEO | 2012/06/01

SECONDARY ACCESSION(S): PRJNA149197

REPOSITORIES: GEO

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Protein Kinase C Mediated Extraembryonic Endoderm Differentiation of Human Embryonic Stem Cells

Project description:Unlike mouse embryonic stem (ES) cells, which are closely related to the inner cell mass, human ES cells appear to be more closely related to the later primitive ectoderm. For example, human ES cells and primitive ectoderm share a common epithelial morphology, growth factor requirements, and the potential to differentiate to all three embryonic germ layers. However, it has previously been shown that human ES cells can also differentiate to cells expressing markers of trophoblast, an extraembryonic lineage formed before the formation of primitive ectoderm. Here we show that phorbol ester 12-O-Tetradecanoylphorbol 13-acetate (TPA) causes human ES cells to undergo an epithelial mesenchymal transition and to differentiate into cells expressing markers of parietal endoderm, another extraembryonic lineage. We further confirmed that this differentiation is through the activation of PKC pathway and demonstrated that a particular PKC subtype, PKC-delta, is most responsible for this transition. This is a time course design. It includes 16 samples.

2012-05-31 | E-GEOD-33180 | biostudies-arrayexpress

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Hypoxia induces a transcriptional primitive streak signature in ES cells and enhances lineage patterning in gastruloids

Project description:The cellular microenvironment shapes stem cell identity and differentiation capacity. Mammalian early embryos are exposed to hypoxia in vivo and benefit from hypoxic culture in vitro. Yet, how different components of the hypoxia response impact stem cell transcriptional networks and lineage choices remains unclear. Here we investigated the effect of acute and prolonged hypoxia on stem cell states and differentiation efficiencies of embryonic and extraembryonic cells. We show that prolonged hypoxia enhances differentiation of embryonic stem (ES) cells towards the mesendoderm lineage by transcriptionally priming cells with a primitive streak signature including Wnt3 and T expression. Exposure to hypoxia in ES culture or during formation of gastrulation-mimicking organoids (gastruloids) moderates T expression and enhances structural complexity. Hypoxic gastruloids generated without exogenous Wnt induction can spontaneously elongate and self-organize. Direct gene regulation by Hif1a, combined with DNA demethylation and metabolic rewiring modulate the transcriptional response and phenotypic outcome. Our findings highlight the influence of the microenvironment on stem cell function and provide a rationale supportive of applying physiological conditions in synthetic embryo models.

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