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Efficient generation of marmoset primordial germ cell-like cells using induced pluripotent stem cells

ABSTRACT: Efficient generation of marmoset primordial germ cell-like cells using induced pluripotent stem cells

PROVIDER: PRJNA866276 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR20680256_1.fastq.gz	Fastqsanger.gz
	SRR20680256_2.fastq.gz	Fastqsanger.gz
	SRR20680257_1.fastq.gz	Fastqsanger.gz
	SRR20680257_2.fastq.gz	Fastqsanger.gz
	SRR20680258_1.fastq.gz	Fastqsanger.gz

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Efficient generation of marmoset primordial germ cell-like cells using induced pluripotent stem cells [scRNA-seq]

Project description:Reconstitution of germ cell fate from pluripotent stem cells provide the opportunities to understand the molecular underpinning of germ cell development. Here, we established a several culture methods of induced pluripotent stem cells (iPSCs) in common marmoset (Callithrix jacchus, cj), which stably propagate in an undifferentiated state. Notably, iPSCs cultured on feeder layer in the presence of a WNT signaling inhibitor upregulate genes related to ubiquitin-dependent protein catabolic process and acquire permissive state that readily differentiate into primordial germ cell-like cells (PGCLCs) bearing immunophenotypic and transcriptomic similarities to pre-migratory cjPGCs. Induction of cjPGCLCs accompanies transient upregulation of mesodermal genes and culminate in establishing primate-specific transcriptional network similar to humans and cynomolgus monkeys. Moreover, cjPGCLCs can be expanded in monolayer while retaining cellular state. Upon co-culture with mouse testicular somatic cells, these cells acquire DDX4+ early prospermatogonia-like state. Our findings provide a framework for understanding and reconstituting germ cell development in marmoset in vitro, which serve as an comparative tool and foundation for a preclinical model for human in vitro gametogenesis.

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Efficient generation of marmoset primordial germ cell-like cells using induced pluripotent stem cells [bisulfite-seq]

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Efficient generation of marmoset primordial germ cell-like cells using induced pluripotent stem cells [bulk RNA-seq]

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Efficient generation of marmoset primordial germ cell-like cells using induced pluripotent stem cells [bisulfite-seq]

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Efficient generation of marmoset primordial germ cell-like cells using induced pluripotent stem cells [scRNA-seq]

Project description:Efficient generation of marmoset primordial germ cell-like cells using induced pluripotent stem cells [scRNA-seq]

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Efficient generation of marmoset primordial germ cell-like cells using induced pluripotent stem cells [bulk RNA-seq]

Project description:Efficient generation of marmoset primordial germ cell-like cells using induced pluripotent stem cells [bulk RNA-seq]

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Generation of marmoset primordial germ cell-like cells under chemically defined conditions

Project description:Primordial germ cells (PGCs) are the embryonic precursors of sperm and oocytes and are responsible for the transmission of genetic and epigenetic information across generations. The development of mouse gametes from PGCs can be reconstituted from pluripotent stem cells in vitro, opening up new avenues for the study of germ cell biology in biomedical research. However, the development of germ cells exhibits molecular and morphological differences between rodents and humans. Therefore, to establish an in vitro model system that is closely related to humans, we studied PGC development in vivo and in vitro in the common marmoset Callithrix jacchus (cj), a New World monkey. Post-migratory cjPGCs in gonads at embryonic day 74 express the germ cell-related genes SOX17, AP2Ɣ, BLIMP1 and the pluripotency markers NANOG and OCT4A (POU5F1), which is reminiscent of human PGCs. We established transgene-free induced pluripotent stem cell (cjiPSC) lines from foetal and postnatal fibroblasts. These cjiPSCs, cultured in a defined medium and in feeder-free conditions, can be differentiated into precursors of mesendoderm and subsequently into cjPGC-like cells (cjPGCLCs) with a transcriptome similar to human PGCs/PGCLCs. Our results not only pave the way to study PGC development in a non-human primate in vitro under experimentally controlled conditions without foetal calf serum and feeder cells, but also provide the opportunity to derive functional marmoset gametes in vitro in future studies.

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Project description:Reconstitution of germ cell lineage using pluripotent stem cells provides a unique platform to deepen our understanding of the mechanisms underlying germ cell development and to produce functional gametes for reproduction. Here, we report a robust induction of primordial germ cell-like cells (PGCLCs) from common marmoset (Callithrix jacchus) embryonic stem cells. The robust induction was achieved by not only activation of the conserved PGC-inducing signals, WNT and BMP4, but also temporal inhibitions of WNT and retinoic acid signals, which prevent mesodermal and neural differentiation, respectively, during PGCLC differentiation. Many of the gene expression and differentiation properties of common marmoset PGCLCs were similar to those of human PGCLCs, making this culture system a reliable and useful primate model. Finally, we identified PDPN and KIT as surface marker proteins by which PGCLCs can be isolated from embryonic stem cells without genetic manipulation. This study will expand the opportunities for research on germ cell development and production of functional gametes to the common marmoset.

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