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Transcriptome analysis reveals conditions for culturing human primitive undifferentiated spermatogonia

ABSTRACT: Transcriptome analysis reveals conditions for culturing human primitive undifferentiated spermatogonia

PROVIDER: PRJNA602712 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR10953067.fastq.gz	Fastqsanger.gz
	SRR10953068.fastq.gz	Fastqsanger.gz
	SRR10953069.fastq.gz	Fastqsanger.gz
	SRR10953070.fastq.gz	Fastqsanger.gz
	SRR10953071.fastq.gz	Fastqsanger.gz

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Transcriptome analysis reveals conditions for culturing human primitive undifferentiated spermatogonia

Project description:Spermatogonial stem cells (SSCs) are essential for the generation of sperm and have potential therapeutic value for male infertility, which afflicts >100 million men world-wide. To devise SSC therapy approaches, it is critical to first develop methods to culture human SSCs in vitro. Here, we report on a transcriptome approach to address this question. Using single-cell RNA-seq (scRNAseq), immunofluorescence, and germ-cell xenograft transplantation analyses, we identified a cell-surface protein, PLPPR3, that purifies human primitive undifferentiated spermatogonia (uSPG) enriched for SSCs. Comparative RNAseq analysis of PLPPR3+ cells (primitive uSPG) with KIT+ cells (enriched for differentiating [d] SPG) revealed that these two stages differentially express a remarkably large number of genes, including genes encoding key components in the TGF, GDNF, AKT, and JAK-STAT signaling pathways. Using scRNAseq analysis and conventional approaches, we tested the effect of manipulating these signaling pathways on cultured human SPG. This revealed that GDNF and BMP8B broadly support the culture of SPG, Activin A supports more advanced SPG, and one condition—AKT pathway inhibition—had the unique ability to selectively support primitive uSPG. These findings have implications for methods to culture and expand human SSCs for therapeutic uses in the future.

2020-06-29 | GSE144085 | GEO

DAZL targets in undifferentiated spermatogonia

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Project description:The maintenance of spermatogenesis in adult males is dependent on a population of mitotic germ cells with self-renewal potential known as undifferentiated spermatogonia. Regulation of undifferentiated spermatogonia function is dependent on transcriptional and post-transcriptional mechanisms. We have identified an essential role for the RNA helicase DDX5 in undifferentiated spermatogonia through generation of a UBC-CreERT2;Ddx5flox/flox mouse model that allows tamoxifen-dependent Ddx5 ablation. To identify genes regulated by DDX5, we generated lines of cultured undifferentiated spermatogonia from these mice and treated cells with tamoxifen (TMX) to induce Ddx5 knockout or vehicle (VEH) as a control, and performed RNA-sequencing analysis to compare these conditions.

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2023-12-30 | E-MTAB-11777 | biostudies-arrayexpress

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