Robust self-organization of livestock pluripotent stem cells into post-gastrulation embryo models with advanced neuronal and mesodermal structures

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The study aimed to overcome limitations of stem cell–based embryo models by creating post-gastrulation models in ungulate species, specifically sheep and pig, using pluripotent stem cell–derived aggregates. The authors generated gastruloids that recapitulate gastrulation features such as germ layer specification, symmetry breaking, and axial elongation, and developed ovine trunk-like structures that robustly model post-gastrulation trunk development with sustained elongation, neuromesodermal progenitor maintenance, segmented somite formation, and a central neural tube–like axis. Time-resolved single-cell RNA sequencing with immunostaining showed coordinated emergence of neural, mesodermal, and intermediate mesodermal lineages arranged along an anteroposterior axis, and the trunk models produced dorsal neural derivatives, anterior neuronal populations, and renal primordia, expanding lineage repertoire beyond prior trunk models. A limitation noted by the authors is that some figure legends next to UMAP plots and supplementary figure legends were corrupted during export but re-uploaded without other content changes. The paper does not explicitly discuss endometriosis or adenomyosis; it was included in the corpus via a keyword match in the upstream search index.

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Abstract

Mammalian body plan formation arises from the self-organization of pluripotent cells through conserved morphogenetic processes that are difficult to study in vivo. Stem cell–based embryo models (SEMs) offer accessible three-dimensional systems to investigate these events but are currently limited to mouse and human cells and largely recapitulate posterior embryonic structures. In addition, no in-vitro models exist for post-gastrulation development in ungulate species, whose early development differs from that of rodents and primates. Here, we establish SEMs for two common ungulates, sheep and pig, using pluripotent stem cell–derived aggregates. We generate ovine and porcine gastruloids that recapitulate key features of gastrulation, including germ layer specification, symmetry breaking, and axial elongation. We further develop ovine trunk-like structures (oTLSs) that robustly model post-gastrulation trunk development, exhibiting sustained elongation, neuromesodermal progenitor maintenance, segmented somite formation, and a central neural tube–like axis. Time-resolved single-cell RNA sequencing combined with immunostaining reveals coordinated emergence of neural, mesodermal, and intermediate mesodermal lineages arranged along an anteroposterior axis. Notably, oTLSs generate dorsal neural derivatives, anterior neuronal populations, and renal primordia, representing an expansion in the lineage repertoire reported for existing trunk models. Together, this work extends SEMs to livestock species and establishes a platform for comparative mammalian developmental studies, with potential applications in fundamental research, veterinary toxicology, and agricultural biotechnology.
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Abstract Mammalian body plan formation arises from the self-organization of pluripotent cells through conserved morphogenetic processes that are difficult to study in vivo. Stem cell–based embryo models (SEMs) offer accessible three-dimensional systems to investigate these events but are currently limited to mouse and human cells and largely recapitulate posterior embryonic structures. In addition, no in-vitro models exist for post-gastrulation development in ungulate species, whose early development differs from that of rodents and primates. Here, we establish SEMs for two common ungulates, sheep and pig, using pluripotent stem cell–derived aggregates. We generate ovine and porcine gastruloids that recapitulate key features of gastrulation, including germ layer specification, symmetry breaking, and axial elongation. We further develop ovine trunk-like structures (oTLSs) that robustly model post-gastrulation trunk development, exhibiting sustained elongation, neuromesodermal progenitor maintenance, segmented somite formation, and a central neural tube–like axis. Time-resolved single-cell RNA sequencing combined with immunostaining reveals coordinated emergence of neural, mesodermal, and intermediate mesodermal lineages arranged along an anteroposterior axis. Notably, oTLSs generate dorsal neural derivatives, anterior neuronal populations, and renal primordia, representing an expansion in the lineage repertoire reported for existing trunk models. Together, this work extends SEMs to livestock species and establishes a platform for comparative mammalian developmental studies, with potential applications in fundamental research, veterinary toxicology, and agricultural biotechnology. Competing Interest Statement The authors have declared no competing interest. Footnotes Some legends next to UMAP plots were corrupted during export, in Figures 1 and 3, and some of the Supp figures. Newly exported versions were uploaded. No other change to the content of the text or figures.

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last seen: 2026-05-20T01:45:00.602351+00:00