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Gastruloid-derived Primordial Germ Cell-like Cells (Gld-PGCLCs) develop dynamically within integrated tissues

ABSTRACT: Gastruloid-derived Primordial Germ Cell-like Cells (Gld-PGCLCs) develop dynamically within integrated tissues

PROVIDER: PRJNA949642 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR23997132_1.fastq.gz	Fastqsanger.gz
	SRR23997132_2.fastq.gz	Fastqsanger.gz
	SRR23997133_1.fastq.gz	Fastqsanger.gz
	SRR23997133_2.fastq.gz	Fastqsanger.gz
	SRR23997134_1.fastq.gz	Fastqsanger.gz

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Gastruloid-derived Primordial Germ Cell-like Cells (Gld-PGCLCs) develop dynamically within integrated tissues

Project description:Primordial Germ Cells (PGCs) are a rare population of cells specified early in embryonic development that give rise to gametes - sperm and egg cells. Current in vitro systems to derive PGC-like cells (PGCLCs) from pluripotent cells use large embryoid bodies, exposed to signalling cocktails, that do not recapitulate the native embryonic environment during PGC formation. Here we show that mouse gastruloids, three-dimensional in vitro models of gastrulation, generate a population of Gastruloid-derived PGC-like cells (Gld-PGCLCs) that resemble early PGCs in vivo. Importantly, the conserved organisation of mouse gastruloids leads to coordinated spatial and temporal localisation of Gld-PGCLCs relative to surrounding somatic cells, even in the absence of exogenous PGC-specific signalling or extraembryonic tissues. In gastruloids, self-organised interactions between cells and tissues, including the endodermal epithelium, enables the specification, migration, and subsequent maturation of a pool of Gld-PGCLCs. As such, mouse gastruloids represent a new source of PGCLCs in vitro and, due to their inherent co-development, serve as a novel model of dynamic PGC development within integrated tissue environments.

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Project description:Gastruloids are highly scalable, three-dimensional assemblies generated from pluripotent stem cells that recapitulate fundamental principles of embryonic pattern formation in vitro. Using single cell RNA and multiome sequencing we provide a comprehensive resource mapping cellular states and cell types found during gastruloid development and compare them to the in vivo embryo. We further develop a high throughput gastruloid handling and imaging pipeline to spatially monitor cell type emergence and unfolding of symmetry breaking during gastruloid development. We report spatial variability of pluripotency states in early gastruloids that determines a binary cell response to Wnt activation. While cells situated in the core of the gastruloid revert to an ectopic pluripotent state, peripheral cells differentiate to a primitive streak like state. These two populations then cause gastruloids to break radial symmetry, allowing axial elongation and commitment to the three embryonic germ layers. Finally by performing a phenotypic compound screen, we perturb thousands of gastruloids at relevant developmental time points deriving a phenotypic landscape and inferring molecular regulator networks underlying gastruloid development. Employing this resource, we improve the formation of anterior structures in the existing gastruloid model, using a dual Wnt modulation approach to differentiate an anterior ectopic pluripotent core to anterior ecto- and endodermal structures. This work gives is a resource to understand how gastruloids develop and, more generally, how homogenous cell populations can generate complex patterns in vitro.

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Project description:We report the Tex10 binding and genome-wide map of H3K4me3 modification in Day 2 PGCLCs by chromatin immunoprecipitation sequencing.

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Project description:Reconstitution of female germ-cell development in vitro is a key challenge in reproductive biology and medicine. We show here that female (XX) embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) in mice are induced into primordial germ cell-like cells (PGCLCs), which, when aggregated with female gonadal somatic cells as reconstituted ovaries, develop pre-meiotic germ cell characteristics, including X-reactivation, imprint erasure, and cyst formation. Upon transplantation under ovarian bursa, PGCLCs in the reconstituted ovaries mature into germinal vesicle-stage oocytes, which, through in vitro maturation and fertilization, contribute to fertile offspring. Our culture system serves as a robust foundation for the investigation of key properties of female germ cells, including the acquisition of totipotency, and for the reconstitution of whole female germ-cell development in vitro. We performed expression analysis of female PGC-like cells [PGCLCs; day 3 (d3), day 6 (d6), and day 3 followed by day 6 in aggregation with female embryonic day (E) 12.5 gonadal somatic cells (d3ag6)] in comparison to in vivo embryonic PGCs (E12.5 female PGCs and E9.5 PGCs) and male d6 PGCLCs. PGCLCs were marked with fluorescence of Blimp1-mVenus, stella-ECFP transgenic reporters (BVSC). PGCs were marked with fluorescence of the stella-EGFP transgenic reporter.

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Project description:We developed a mRNA transfection-based method for the efficient generation of primordial germ cell like cells (PGCLCs) in marmoset. Single cell RNA-seq analyses (10X) confirmed that the induced marmoset PGCLCs show transcriptome profile similar to in vivo PGCs. For comparison we sequenced iPSCs, E74 ovaries, E82 ovaries, NB ovaries, E87 testes, and day 22 testes.

2023-08-11 | E-MTAB-12123 | biostudies-arrayexpress

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