BMP4 signaling directs primitive endoderm-derived XEN cells to an extraembryonic visceral endoderm identity

Artus, Jérôme; Douvaras, Panagiotis; Piliszek, Anna; Isern, Joan; Baron, Margaret H.; Hadjantonakis, Anna-Katerina

doi:10.1016/j.ydbio.2011.10.015

Cited by 72 publications

(89 citation statements)

References 99 publications

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“…S6). The expression of endoderm transcription factors is not upregulated compared with untreated controls, with the exception of Gata4, which is over twofold enriched, confirming previous observations (Artus et al, 2012). Taken together, this demonstrates that cXEN cells, like embryo-derived XEN cells, have the capacity to differentiate into VE cells in response to BMP4 treatment.…”

Section: Research Articlesupporting

confidence: 88%

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Conversion from mouse embryonic to extra-embryonic endoderm stem cells reveals distinct differentiation capacities of pluripotent stem cell states

et al. 2012

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SUMMARYThe inner cell mass of the mouse pre-implantation blastocyst comprises epiblast progenitor and primitive endoderm cells of which cognate embryonic (mESCs) or extra-embryonic (XEN) stem cell lines can be derived. Importantly, each stem cell type retains the defining properties and lineage restriction of their in vivo tissue of origin. Recently, we demonstrated that XEN-like cells arise within mESC cultures. This raises the possibility that mESCs can generate self-renewing XEN cells without the requirement for gene manipulation. We have developed a novel approach to convert mESCs to XEN cells (cXEN) using growth factors. We confirm that the downregulation of the pluripotency transcription factor Nanog and the expression of primitive endoderm-associated genes Gata6, Gata4, Sox17 and Pdgfra are necessary for cXEN cell derivation. This approach highlights an important function for Fgf4 in cXEN cell derivation. Paracrine FGF signalling compensates for the loss of endogenous Fgf4, which is necessary to exit mESC selfrenewal, but not for XEN cell maintenance. Our cXEN protocol also reveals that distinct pluripotent stem cells respond uniquely to differentiation promoting signals. cXEN cells can be derived from mESCs cultured with Erk and Gsk3 inhibitors (2i), and LIF, similar to conventional mESCs. However, we find that epiblast stem cells (EpiSCs) derived from the post-implantation embryo are refractory to cXEN cell establishment, consistent with the hypothesis that EpiSCs represent a pluripotent state distinct from mESCs. In all, these findings suggest that the potential of mESCs includes the capacity to give rise to both extra-embryonic and embryonic lineages. KEY WORDS: Pluripotent stem cells, Directed differentiation, Extra-embryonic endoderm, FGF, Mouse embryoConversion from mouse embryonic to extra-embryonic endoderm stem cells reveals distinct differentiation capacities of pluripotent stem cell states RESEARCH ARTICLE Conversion of mES to cXEN cellsNevertheless, it remains unclear whether self-renewing XEN cells can be derived directly from mESCs without requiring transgenic over-expression.The fibroblast growth factor (FGF) receptor Fgfr2 is enriched in PrE cells, and the ligand Fgf4 is expressed by epiblast progenitor cells within the ICM (Feldman et al., 1995;Arman et al., 1998; Guo et al., 2010). This complementary receptor-ligand expression suggests that epiblast-secreted Fgf4 may be functionally important for PrE development (Rappolee et al., 1994; Goldin and Papaioannou, 2003). It has recently been suggested that PrE formation requires non-cell-autonomous provision of Fgf4 by Nanog-expressing cells (Nichols et al., 2009;Messerschmidt and Kemler, 2010;Yamanaka et al., 2010; Frankenberg et al., 2011). Indeed, Fgf4 or Fgf2, which are not expressed in the early embryo, both function via Fgfr2 and are routinely added during XEN derivation from embryos (Kunath et al., 2005). However, it is unclear whether FGFs are required for XEN cell derivation or whether they function in an autocrine or paracrine m...

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Section: Research Articlesupporting

confidence: 88%

“…Recently, a subtype of VE cells has been differentiated from embryo-derived XEN cells treated with BMP4, demonstrating the plasticity of XEN cells (Paca et al, 2012;Artus et al, 2012 Fig. S5).…”

Section: Cxen Cells Undergo Ve Differentiation In Response To Bmp Trementioning

confidence: 96%

Conversion from mouse embryonic to extra-embryonic endoderm stem cells reveals distinct differentiation capacities of pluripotent stem cell states

et al. 2012

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“…Recent studies on mouse XEN cells demonstrated that Nodal/ Cripto signaling induced the anterior VE formation ( Kruithof-de Julio et al, 2011), whereas BMP4 promoted the extraembryonic VE differentiation (Artus et al, 2012;Paca et al, 2012). We found that treatment with Nodal did not cause any morphological transition in rat XEN cells (data not shown).…”

Section: Formation and Characterization Of Tight Junctions In The Presupporting

confidence: 52%

“…We found that treatment with Nodal did not cause any morphological transition in rat XEN cells (data not shown). However, when we treated the cells growing on laminin in N2B27 medium with BMP4 following the established protocol (Artus et al, 2012;Paca et al, 2012) (Fig. 5A), we noted the formation of an epithelium on day 2 (1 day after the BMP4 treatment) (Fig.…”

Section: Formation and Characterization Of Tight Junctions In The Prementioning

confidence: 97%

“…Nevertheless, differentiation to VE cells, specifically anterior VE cells, has been achieved with mouse XEN cells using the ligands of Nodal and Cripto pathways (Kruithof-de Julio et al, 2011). More recently, mouse XEN cells were differentiated into extraembryonic VE cells after treatment with BMP ligands (Artus et al, 2012;Paca et al, 2012).…”

Section: Introductionmentioning

confidence: 99%

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Activation of the PTHRP/adenylate cyclase pathway promotes differentiation of rat XEN cells into parietal endoderm, whereas Wnt/β-catenin signaling promotes differentiation into visceral endoderm

Chuykin

Schulz

Guan

et al. 2013

Journal of Cell Science

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SummaryDuring early mammalian development, primitive endoderm (PrE) is specified and segregated away from the pluripotent epiblast. At a later developmental stage, PrE forms motile parietal endoderm (PE) lying proximal to the trophectoderm, and visceral endoderm (VE) that contacts the developing epiblast and extraembryonic ectoderm. Mouse extraembryonic endoderm (XEN) cells were isolated and became widely used to study signals governing lineage specification. Rat XEN cell lines have also been derived, but were distinguished from mouse by expression of SSEA1 and Oct4. We showed here that rat XEN cells grown in the presence of a GSK3 inhibitor or overexpressing b-catenin exhibited enhanced formation of cell contacts and decreased motility. Rat XEN cells treated with BMP4 revealed similar morphological changes. Furthermore, we observed that rat XEN cells cultured with GSK3 inhibitor formed adhesion and tight junctions, and acquired bottom-top polarity, indicating the formation of VE cells. In contrast, forskolin, an activator of the cAMP pathway, induced the disruption of cell contacts in rat XEN cells. Treatment with forskolin induced PE formation and epithelialmesenchymal transition (EMT) in rat XEN cells. Using microarray and real-time PCR assays, we found that VE versus PE formation of rat XEN cells was correlated with change in expression levels of VE or PE marker genes. Similar to forskolin, EMT was prompted upon treatment of rat XEN cells with recombinant parathyroid hormone related peptide (PTHRP), an activator of the cAMP pathway in vivo. Taken together, our data suggest that rat XEN cells are PrE-like cells. The activation of Wnt or BMP4 pathways in rat XEN cells leads to the acquisition of VE characteristics, whereas the activation of the PTHRP/cAMP pathway leads to EMT and the formation of PE.

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Automated, High‐Throughput Phenotypic Screening and Analysis Platform to Study Pre‐ and Post‐Implantation Morphogenesis in Stem Cell‐Derived Embryo‐Like Structures

Shankar,

van Blitterswijk,

Vrij

et al. 2023

Advanced Science

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Combining high‐throughput generation and high‐content imaging of embryo models will enable large‐scale screening assays in the fields of (embryo) toxicity, drug development, embryogenesis, and reproductive medicine. This study shows the continuous culture and in situ (i.e., in microwell) imaging‐based readout of a 3D stem cell‐based model of peri‐implantation epiblast (Epi)/extraembryonic endoderm (XEn) development with an expanded pro‐amniotic cavity (PAC) (E3.5 E5.5), namely XEn/EPiCs. Automated image analysis and supervised machine learning permit the identification of embryonic morphogenesis, tissue compartmentalization, cell differentiation, and consecutive classification. Screens with signaling pathway modulators at different time windows provide spatiotemporal information on their phenotypic effect on developmental processes leading to the formation of XEn/EPiCs. Exposure of the biological model in the microwell platform to pathway modulators at two time windows, namely 0–72 h and 48–120 h, show that Wnt and Fgf/MAPK pathway modulators affect Epi differentiation and its polarization, while modulation of BMP and Tgfβ/Nodal pathway affects XEn specification and epithelialization. Further, their collective role is identified in the timing of the formation and expansion of PAC. The newly developed, scalable culture and analysis platform, thereby, provides a unique opportunity to quantitatively and systematically study effects of pathway modulators on early embryonic development.

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BMP4 signaling directs primitive endoderm-derived XEN cells to an extraembryonic visceral endoderm identity

Cited by 72 publications

References 99 publications

Conversion from mouse embryonic to extra-embryonic endoderm stem cells reveals distinct differentiation capacities of pluripotent stem cell states

Conversion from mouse embryonic to extra-embryonic endoderm stem cells reveals distinct differentiation capacities of pluripotent stem cell states

Activation of the PTHRP/adenylate cyclase pathway promotes differentiation of rat XEN cells into parietal endoderm, whereas Wnt/β-catenin signaling promotes differentiation into visceral endoderm

Automated, High‐Throughput Phenotypic Screening and Analysis Platform to Study Pre‐ and Post‐Implantation Morphogenesis in Stem Cell‐Derived Embryo‐Like Structures

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