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Single-cell RNA-sequencing of mouse embryoids at Day 8

ABSTRACT: Generating properly organized and differentiated embryonic structures in vitro from aggregates of pluripotent cells remains a major challenge to overcome. In the living embryo, the interplay between morphogens and their antagonists set up gradients of activity that instruct cells about their fate, leading to patterning and morphogenesis. Here we show that experimentally engineering a morphogen signaling center within an aggregate of mouse pluripotent stem cells is sufficient to mimic in vitro the function of an embryo organizer and to trigger embryonic development. The resulting embryoids are extensively patterned along their anterior-posterior and dorsal-ventral axes, with formation of three germ layers through a process of gastrulation and differentiation of germ layer derivatives similar to those of a neurula-stage mouse embryo.

ORGANISM(S): Mus musculus

PROVIDER: GSE168539 | GEO | 2021/04/22

REPOSITORIES: GEO

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Construction of a Mammalian Embryo from Embryonic Stem Cells Instructed by Morphogen Gradients

Project description:Generating properly organized and differentiated embryonic structures in vitro from aggregates of pluripotent cells remains a major challenge. In a living embryo, the interplay in between signaling molecules known as morphogens and their antagonists lead to gradients of activity that instruct cells about their fate, leading to patterning and morphogenesis. Here we show that experimentally engineering a morphogen signaling center within an aggregate of mouse pluripotent stem cells is sufficient to mimic the function of an embryo organizer and to trigger embryonic development. The resulting embryoids are remarkably well patterned along anterior-posterior and dorsal-ventral axes, generate three germ layers through a process of gastrulation and differentiate germ layer derivatives (including notochord, vasculature, neural tube, neural crest, endoderm) similar to an authentic embryo at E9-E9.5.

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Single-cell RNA sequencing data of stem cell-derived microfluidic human embryoids, NODAL-KO human embryoids and chimpanzee embryoids

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2021-10-13 | GSE185643 | GEO

Mouse complete embryoid development within a yolk sac recapitulates gastrulation, neurulation and early organogenesis

Project description:Embryo-like structures generated from stem cells can achieve varying developmental milestones, but none have been shown to progress through gastrulation, neurulation, and organogenesis. Here, we show that "ETiX" mouse embryoids, assembled from embryonic stem cells, trophoblast stem cells and inducible extraembryonic endoderm stem cells, can develop into gastrulating embryoids, and beyond to generate neurulating embryoids, which generate the progenitors needed to create the entire organism. The head-folds of ETiX neurulating embryoids show anterior expression of Otx2, defining forebrain and midbrain regions that resemble those of the natural mouse embryo. Neurulating embryoids also develop beating heart-like structures, trunks comprising a neural tube and somites, tail buds containing neuromesodermal progenitors and primordial germ cells, and gut tubes derived from definitive endoderm. Notably, neurulating embryoids also develop a yolk sac with blood islands. Overall, ETiX neurulating embryoid formation strongly resembles natural embryogenesis, advancing embryo-like development further than any other stem-cell derived model and within extra-embryonic membranes.

2022-08-12 | GSE209792 | GEO

Single-cell RNA sequencing data of stem cell-derived microfluidic human embryoids, NODAL-KO human embryoids and chimpanzee embryoids

Project description:Single-cell RNA sequencing data of stem cell-derived microfluidic human embryoids, NODAL-KO human embryoids and chimpanzee embryoids

| PRJNA770264 | ENA

Retinoic Acid Delivery within Embryoid Bodies Induces an Early Streak Phenotype in vitro

2009-08-04 | GSE12333 | GEO

Efficient generation of ETX embryoids that recapitulate the entire window of murine egg cylinder development

Project description:The murine embryonic–trophoblast–extra-embryonic endoderm (ETX) model is an integrated stem cell–based model to study early postimplantation development. It is based on the self-assembly potential of embryonic, trophoblast, and hypoblast/primitive/visceral endoderm-type stem cell lines (ESC, TSC, and XEN, respectively) to arrange into postimplantation egg cylinder–like embryoids. Here, we provide an optimized method for reliable and efficient generation of ETX embryoids that develop into late gastrulation in static culture conditions. It is based on transgenic Gata6-overproducing ESCs and modified assembly and culture conditions. Using this method, up to 43% of assembled ETX embryoids exhibited a correct spatial distribution of the three stem cell derivatives at day 4 of culture. Of those, 40% progressed into ETX embryoids that both transcriptionally and morphologically faithfully mimicked in vivo postimplantation mouse development between E5.5 and E7.5. The ETX model system offers the opportunity to study the murine postimplantation egg cylinder stages and could serve as a source of various cell lineage precursors.

2023-01-25 | GSE206132 | GEO

Epithelial Disruption Drives Mesendoderm Differentiation in Human Pluripotent Stem Cells by Enabling TGF-β Protein Sensing

Project description:The processes of primitive streak formation and fate specification in the mammalian epiblast rely on complex interactions between morphogens and tissue organization. Little is known about how these instructive cues functionally interact to regulate gastrulation. We interrogated the interplay between tissue organization and morphogens by using human induced pluripotent stem cells (hiPSCs) down-regulated for the morphogen regulator GLYPICAN-4, in which defects in tight junctions result in areas of disrupted epithelial integrity. Remarkably, this phenotype does not affect hiPSC stemness, but impacts on cell fate acquisition. Strikingly, cells within disrupted areas become competent to perceive the gastrulation signals BMP4 and ACTIVIN A, (an in vitro surrogate for NODAL), and thus differentiate into mesendoderm. Yet, disruption of epithelial integrity sustains activation of BMP4 and ACTIVIN A downstream effectors and correlates with enhanced hiPSC endoderm/mesoderm differentiation. Altogether, our results disclose epithelial integrity as a key determinant of TGF-β activity and highlight an additional mechanism guiding morphogen sensing and spatial cell fate change within an epithelium.

2023-01-09 | GSE222186 | GEO

Bioengineered embryoids mimic post-implantation development in vitro [EmbryoidsTruseq3]

Project description:Here, we report a bioengineering-inspired approach to simultaneously coax mouse embryonic stem cells (ESCs) into hundreds of uniform epiblast-like (EPI) aggregates in a solid matrix-free manner. When co-cultured with mouse trophoblast stem cell (TSC) aggregates, the resulting hybrid structures initiate gastrulation-like events and undergo axial morphogenesis to yield structures, termed EpiTS embryoids, with a pronounced anterior development, including brain-like regions.

2021-04-12 | GSE171211 | GEO

Bioengineered embryoids mimic post-implantation development in vitro [Embryoids10XG]

2021-04-12 | GSE171209 | GEO

Transcription profiling of mouse embryoid bodies treated with retinoic acid delivery induces an early streak phenotype in vitro

Project description:During embryogenesis, cell specification and tissue formation is directed by the concentration and temporal presentation of morphogens, and similarly, pluripotent embryonic stem cells differentiate in vitro into various phenotypes in response to morphogen treatment. Embryonic stem cells are commonly differentiated as three dimensional spheroids called embryoid bodies (EBs); however, differentiation within EBs is typically heterogeneous and disordered. Here we show that spatiotemporal control of microenvironmental cues embedded directly within EBs enhances the homogeneity, synchrony and organization of differentiation. Degradable polymer microspheres releasing retinoic acid within EBs induce the formation of cystic spheroids closely resembling the early streak mouse embryo, with an exterior of visceral endoderm enveloping an epiblast layer. These results demonstrate that controlled morphogen presentation to stem cells more efficiently directs cell differentiation and tissue formation, thereby improving developmental biology models and enabling the development of regenerative medicine therapies and cell diagnostics. Experiment Overall Design: ESCs (D3 line) were maintained in an undifferentiated state on gelatin-coated tissue culture plates in LIF-containing media as described previously. EBs were formed from 2x106 ESCs inoculated in LIF-free media and cultured under rotary conditions. EBs containing microspheres were produced by coating CellTracker Red (CONTROLs) or RA-loaded (EXPERIMENTs) microspheres in 0.1% gelatin solution for 3 hours prior to mixing with ESCs in various microsphere to cell ratios and a range of rotary speeds. EB media was exchanged every 1-2 days as needed. Experiment Overall Design: Triplicate control and experiment EBs were processed for microarray.

2009-08-08 | E-GEOD-12333 | biostudies-arrayexpress

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