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Bulk RNA-Seq of human primordial germ cells induced from human induced pluripotent cells

ABSTRACT: Mammalian germ cells first establish as primordial germ cells (PGCs). There is an increasing demand to investigate the characteristics of human germ cells including PGCs. Recent studies have established the methodologies to derive human germ cells such as PGCs from human induced pluripotent cells (iPSCs). The in vitro-derived PGCs are referred to as PGC-like cells (PGCLCs) and exhibit the characteristics of the PGCs immediately after specification. Here we performed bulk RNA-sequencing (RNA-Seq) of PGCLCs and their parental iPSCs to characterize the expression signatures of human genes and transposable elements in PGCLCs in detail.

ORGANISM(S): Homo sapiens

PROVIDER: GSE167570 | GEO | 2022/03/08

REPOSITORIES: GEO

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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. 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.

2012-11-05 | E-GEOD-40716 | biostudies-arrayexpress

Heading towards a dead end: the role of DND1 in germ line differentiation of human iPSCs

Project description:The DND microRNA-mediated repression inhibitor 1 (DND1) is a conserved RNA binding protein (RBP) and plays an important role in survival and maintenance of primordial germ cells (PGCs) and the development of the male germline in zebrafish and mice. It was shown to be expressed in human pluripotent stem cells (PSCs), PGCs, and spermatogonia, but little is known about its specific role in pluripotency and human germline development. Here we use CRISPR/Cas mediated knockout and PGC-like cell (PGCLC) differentiation in human iPSCs to analyse if DND1 (1) plays a role in maintaining pluripotency and (2) in specification of PGCLCs. We generated several clonal lines with biallelic loss of function mutations and analysed their potential to differentiate towards PGCLCs and their gene expression on RNA and protein level via bulk RNA sequencing and mass spectrometry. The generated knockout iPSCs showed no differences in pluripotency gene expression, proliferation nor trilineage differentiation potential, but yielded reduced numbers o PGCLCs compared to their parental iPSCs. RNAseq analysis in PGCLCs showed significantly reduced expression of genes associated with cellular developmental processes and cell differentiation in knockout cells, including known markers for PGCs (NANOS3, SOX17, PRDM1, EPCAM) and naïve pluripotency (TFCP2L, DNMT3L).

2021-12-10 | PXD023802 | Pride

Transcriptomes of mouse PGCLCs isolated from 6-day culture embryoid bodies were compared with transcriptomes of their precur cells (ESCs, iPSCs, and EpiLCs) and E12.5 in vivo mouse PGCs

Project description:Transcriptomes of mouse E12.5 primordial germ cells (PGCs), primordial germ cell-like cells (PGCLCs) isolated from 6-day culture embryoid bodies, and the precursor pluripotent stem cells [embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs)] and epiblast-like cells (EpiLCs) Total RNA was isolated from FACS-enriched, SSEA1+/CD61+ double-positive PGCs and PGCLCs. RNA was also isolated from ESC, iPSC, and EpiLC cultured without enrichment. Transcriptomes were determined using Affymetrix microarray.

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mRNA-based generation of marmoset PGCLCs capable of differentiation into gonocyte-like cells

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

Transcriptomes of mouse PGCLCs isolated from 6-day culture embryoid bodies were compared with transcriptomes of their precur cells (ESCs, iPSCs, and EpiLCs) and E12.5 in vivo mouse PGCs

2016-08-22 | GSE80983 | GEO

Derivation of primodial germ cell-like cells from induced pluripotent stem cells of patients with idiopathic azoospermia

Project description:At present, the etiology of most non-obstructive azoospermia (NOA) remains unclear, and the patients are diagnosed as idiopathic infertility. In vitro generation of patient-specific induced pluripotent stem cells (iPSCs) is an effective approach for exploring the mechanisms of human disease. Here, we established iPSCs from patients with idiopathic NOA, and differentiated them into human germline cells both in vitro and in vivo. Compared with normal hiPSCs, the idiopathic NOA patient-specific iPSCs show less efficiency of primodial germ cell-like cells (PGCLCs) formation in vitro and poor proliferation potential in vivo. Although the transcriptomes of PGCLCs are different from that of embryonic gonadal PGCs, the expression patterns of key genes involved in human PGC specification are generally similar during PGCLC induction process from all iPSCs lines, including the activation of BLIMP1, TFAP2C, NANOS3, SOX17, and T, and the repression of PRDM14 and SOX2. Moreover, the PGCLCs derived from idiopathic NOA patient-specific iPSCs initiate epigenetic reprogramming with repression of 5mC and presence of 5hmC. These findings provide a unique foundation for future mechanism research of male infertility.

2019-02-28 | GSE126009 | GEO

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

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In vitro expansion of mouse primordial germ cell‐like cells recapitulates an epigenetic blank slate [ChIP-Seq]

Project description:The expansion of primordial germ cells (PGCs), the precursors for the oocytes and spermatozoa, is a key challenge in reproductive biology/medicine. Using a chemical screening exploiting PGC‐like cells (PGCLCs) induced from mouse embryonic stem cells (ESCs), we here identify key signaling pathways critical for PGCLC proliferation. We show that the combinatorial application of Forskolin and Rolipram, which stimulate cAMP signaling via different mechanisms, expands PGCLCs up to ~50‐fold in culture. The expanded PGCLCs maintain robust capacity for spermatogenesis, rescuing the fertility of infertile mice. Strikingly, during expansion, PGCLCs comprehensively erase their DNA methylome, including parental imprints, in a manner that precisely recapitulates genome‐wide DNA demethylation in gonadal germ cells, while essentially maintaining their identity as sexually uncommitted PGCs, apparently through appropriate histone modifications. By establishing a paradigm for PGCLC expansion, our system reconstitutes the epigenetic “blank slate” of the germ line, an immediate precursory state for sexually dimorphic differentiation.

2017-05-30 | GSE87645 | GEO

In vitro expansion of mouse primordial germ cell‐like cells recapitulates an epigenetic blank slate [RNA-Seq]

2017-05-30 | GSE87644 | GEO

DNA methylation and hydroxymethylation profiles were compared between mouse PGCLCs isolated from 6-day culture embryoid bodies, their precursor cells (ESCs, iPSCs, and EpiLCs), and E12.5 in vivo mouse PGCs.

Project description:DNA methylation and hydroxymethylation in the genomes of mouse E12.5 primordial germ cells (PGCs), primordial germ cell-like cells (PGCLCs) isolated from 6-day culture embryoid bodies, and the precursor pluripotent stem cells [embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs)] and epiblast-like cells (EpiLCs)

2016-08-22 | GSE81175 | GEO

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