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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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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

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

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

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

Global profiling of DNA methylation erasure in mouse primordial germ cells.

Project description:Epigenetic reprogramming, characterized by loss of cytosine methylation and histone modifications, occurs during mammalian development in primordial germ cells (PGCs), yet the targets and kinetics of this process are poorly characterized. Here we provide a map of cytosine methylation on a large portion of the genome in developing male and female PGCs isolated from mouse embryos. We show that DNA methylation erasure is global and affects genes of various biological functions. We also reveal complex kinetics of demethylation that are initiated at most genes in early PGC precursors around embryonic day 8.0-9.0. In addition, besides intracisternal A-particles (IAPs), we identify rare LTR-ERV1 retroelements and single-copy sequences that resist global methylation erasure in PGCs as well as in preimplantation embryos. Our data provide important insights into the targets and dynamics of DNA methylation reprogramming in mammalian germ cells.

| S-EPMC3317146 | biostudies-literature

NRF1 promotes primordial germ cell development, proliferation and survival [ChIP-seq]

Project description:To investigate the role of NRF1 in regulating primordial germ cell development, We established conditional knockout mice of Nrf1 in primordial germ cell to observe the effect of Nrf1 knockout on the development of primordial germ cell. At the same time, we utilized a pluripotent stem cell differentiation system in vitro to obtain PGCL cells for chip_ Seq, analyze which genes Nrf1 directly binds to. Meanwhile, we established a pluripotent stem cell line induced by Nrf1 overexpression and performed RNA_seq analysis on PGCL cells overexpressing Nrf1 obtained in vitro

2023-08-05 | GSE231661 | GEO

NRF1 promotes primordial germ cell development, proliferation, and survival [RNA-seq]

2023-08-05 | GSE231662 | 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

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

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