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An ExtendedCulture System that Supports Human Primordial Germ Cell-Like Cells Survival and Initiation of DNA Methylation Erasure

ABSTRACT: An ExtendedCulture System that Supports Human Primordial Germ Cell-Like Cells Survival and Initiation of DNA Methylation Erasure

PROVIDER: PRJNA578405 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR10312800.fastq.gz	Fastqsanger.gz
	SRR10312801.fastq.gz	Fastqsanger.gz
	SRR10312802.fastq.gz	Fastqsanger.gz
	SRR10312803.fastq.gz	Fastqsanger.gz
	SRR10312804.fastq.gz	Fastqsanger.gz

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Similar Datasets

Genome-wide erasure of DNA methylation in mouse primordial germ cells is affected by Aid deficiency

Project description:Epigenetic reprogramming including demethylation of DNA occurs in mammalian primordial germ cells (PGCs) and in early embryos, and is important for the erasure of imprints and epimutations, and the return to pluripotency. The extent of this reprogramming and its molecular mechanisms are poorly understood. We previously showed that the cytidine deaminases Aid and Apobec1 can deaminate 5-methylcytosine in vitro and in E coli, and in the mouse are expressed in tissues in which demethylation occurs. Here we profiled DNA methylation throughout the genome by unbiased bisulfite Next Generation Sequencing (BS-Seq) in wildtype and Aid deficient PGCs at E13.5. Wildtype PGCs revealed dramatic genome-wide erasure of methylation to a level below that of methylation deficient (Np95-/-) ES cells, with female PGCs being less methylated than male ones. By contrast, Aid deficient PGCs were up to three times more methylated than wildtype ones; this substantial difference occurred throughout the genome, with introns, intergenic regions and transposons being relatively more methylated than exons. Relative hypermethylation in Aid deficient PGCs was confirmed by analysis of individual loci in the genome. Our results reveal that erasure of DNA methylation in the germ line is a global process, hence limiting the potential for transgenerational epigenetic inheritance. Aid deficiency interferes with genome-wide erasure of DNA methylation patterns, suggesting that Aid has a critical function in epigenetic reprogramming and potentially in restricting the inheritance of epimutations in mammals. Comparison of methylation in wild-type and Aid deficient mouse tissues

2010-01-26 | E-GEOD-19960 | biostudies-arrayexpress

An ExtendedCulture System that Supports Human Primordial Germ Cell-Like Cells Survival and Initiation of DNA Methylation Erasure

Project description:In this study, we evalauted the transcriptionl profiles of human primordial germ cell-like cell (hPGCLCs) cultured for an extended period on STO feeders in FR10 media for an additional 10 days (D4C10). The hPGCLCs were obtained from two different human embryonic stem cell line (hESCs), UCLA 1 (XX) and UCLA2 (XY) for the RNA sequencing data. For whole genome bisulfite sequencing, we evaluate primed UCLA2 hESCs, directly out of the aggragate hPGCLCs (Day 4= D4) and extended culture hPGCLCs D4C10. Our D4C10 hPGCLCs were compared to prior RNA-sequencing data generated in our laboraty, which revealed that D4C10 hPGCLCs retain germline identy and closely resemble D4 hPGCLCs at the transcriptional level. Our WGBS data revealed that extended culture hPGCLCs at D4C10 have a decrease in CG methlyation, as compared to the starting hESCs and D4 hPGCLCs.

2020-01-31 | GSE139115 | GEO

Genome-wide erasure of DNA methylation in mouse primordial germ cells is affected by Aid deficiency

2010-01-26 | GSE19960 | GEO

Genome-wide DNA methylation reprogramming in porcine primordial germ cells

Project description:DNA methylation reprogramming of primordial germ cells (PGCs) is an essential step that affects the activation and inactivation of certain genes, therefore having a direct impact on the transcriptome of an individual. In this study, we have described the methylome landscape of porcine PGCs, characterizing the genomic elements that resist methylation erasure.

2021-02-04 | E-MTAB-9326 | biostudies-arrayexpress

An Extended Culture System that Supports Human Primordial Germ Cell-like Cell Survival and Initiation of DNA Methylation Erasure.

Project description:The development of an in vitro system in which human primordial germ cell-like cells (hPGCLCs) are generated from human pluripotent stem cells (hPSCs) has been invaluable to further our understanding of human primordial germ cell (hPGC) specification. However, the means to evaluate the next fundamental steps in germ cell development have not been well established. In this study we describe a two dimensional extended culture system that promotes proliferation of specified hPGCLCs, without reversion to a pluripotent state. We demonstrate that hPGCLCs in extended culture undergo partial epigenetic reprogramming, mirroring events described in hPGCs in vivo, including a genome-wide reduction in DNA methylation and maintenance of depleted H3K9me2. This extended culture system provides a new approach for expanding the number of hPGCLCs for downstream technologies, including transplantation, molecular screening, or possibly the differentiation of hPGCLCs into gametes by in vitro gametogenesis.

| S-EPMC7066331 | biostudies-literature

The single cell transcriptional landscape of female primordial germ cell differentiation

Project description:Female primordial germ cell differentiation undergoes sex determination and meiosis initiation asynchronously. Here we investigate the transcriptional profiles of 20519 single cells collected from E12.5, E14.5 and E16.5 mouse fetal ovaries. Clustering analysis identifies ten clusters and defines dozens of corresponding marker genes and provides a global view of cellular differentiation from undifferentiated primordial germ cells towards meiotic oocytes. We explore the dynamics of gene expression within the differentiation trajectory with special focus on the mechanisms for meiotic initiation.

2020-04-01 | GSE130212 | GEO

FGFR3 is Expressed by Human Primordial Germ Cells and is Repressed after Meiotic Initiation to Form Primordial Oocytes

Project description:FGFR3 is Expressed by Human Primordial Germ Cells and is Repressed after Meiotic Initiation to Form Primordial Oocytes

| PRJNA796329 | ENA

FGFR3 is Expressed by Human Primordial Germ Cells and is Repressed after Meiotic Initiation to Form Primordial Oocytes

Project description:Human germ cell development is a highly regulated process beginning soon after embryo implantation with the specification of primordial germ cells (PGCs) and ending in adulthood with the differentiation of gametes. Here, we show that fibroblast growth factor receptor 3 (FGFR3) is expressed by human PGCs during the first and second trimester, becoming repressed as PGCs differentiate into primordial oocytes. Using fluorescence activated cell sorting (FACS) with antibodies that recognize FGFR3 followed by single cell RNA sequencing, we show that isolating FGFR3-positive cells enriches for human PGCs. Taken together, FGFR3 could be used in future studies as a strategy to identify maturing hPGCs.

2022-05-19 | GSE193474 | GEO

Heterochromatin Protein 1γ Regulates Epigenetic Reprogramming in Primordial Germ Cells

Project description:Temporal and spatial regulation of trimethylation of histone H3 lysine27 (H3K27me3) is crucial for cell lineage commitment and development. Resetting the ground state of H3K27me3 is important for establishing a proper development program of primordial germ cells (PGCs). The mechanisms that regulate global erasure of epigenetic information in the germ line are not well understood. Here we show that HP1γ suppresses the demethylation activity of Utx through direct interaction in vitro and in vivo. Down-regulation of mammalian heterochromatin protein 1 gamma (HP1γ) coincides with transient loss of H3K27me3 in PGCs on Embryonic Day 10.5 (E10.5). Furthermore, transient abolishment of HP1γ promotes the induction of embryonic stem cells into putative PGCs in vitro. These data reveal a cross-talk between HP1γ and Utx in controlling histone H3K27 methylation status, thereby define HP1γ as a molecular regulator of germ cell epigenetic reprogramming.

2017-01-24 | GSE93924 | GEO

Offspring from oocytes derived from in vitro primordial germ cell-like cells in mice

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.

2012-11-05 | GSE40716 | GEO

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