Browse
Submit Data
Databases
API
Help

Dataset Information

0 Views

0 Connections

0 Citations

0 Reanalyses

0 Downloads

Omics score: 0

Transcriptome analysis reveals conditions for culturing human primitive undifferentiated spermatogonia

ABSTRACT: 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.

ORGANISM(S): Homo sapiens

PROVIDER: GSE144085 | GEO | 2020/06/29

REPOSITORIES: GEO

ACCESS DATA

Json Xml

Dataset's files

Source:

			Action	DRS
		Other

Items per page:

1 - 1 of 1

Similar Datasets

LncRNA Sequencing Analysis of Mouse spermatogonial stem cells during Glial cell-derived neurotrophic factor?GDNF?treatment

Project description:Long non-coding RNAs (lncRNAs) are transcripts longer than 200 nucleotides and have little or no potential for translation. More and more studies have domenstrated mammalian lncRNAs are intrinsically functional and growing data indicate lncRNAs are determinants of stem cell fate by regulating potency, self-renewal and differentiation. There is no report of lncRNAs in spermatogonial stem cells?SSCs?, and importance of lncRNAs in germline linage stem cell has not been investigated yet. here, we presents a large –scale profiling of all lncRNAs in SSCs as well as those lncRNAs regulated by SSC dependent growth factor GDNF through high throughput sequencing. Spermatogonial stem cells were first suffered from an 18 hr GDNF withdrawal followed by refreshment with GDNF for 8 hrs and RNA samples were collected from normal culture wells (N), GDNF 18hr withdrawal wells (0hr), and GDNF 8 hr refreshed cells (8hr). After all GDNF treatment, cultured germ cell clumps were gently blowed with a 200?l pipette, and followed by Total RNA isolation and sequencing by Illumina HiSeqTM 2000

2015-04-15 | E-GEOD-66998 | biostudies-arrayexpress

Neonatal and adult human testis defined at the single-cell level

Project description:Spermatogenesis has been well studied in rodents and invertebrates, but remains poorly understood in humans. As a step towards illuminating human spermatogenesis, we used single-cell RNA-sequencing (scRNAseq) analysis to analyze neonatal and adult human testes. Clustering analysis of neonatal testes revealed 3 germ subsets, including cells with characteristics of primordial germ cells (PGCs), and more differentiated cells with gene expression profiles similar with adult spermatogonial stem cells (SSCs). We identified markers for these neonatal subsets, including protein markers for the PGC-like (PGCL) subset. Clustering analysis of the adult testis revealed 9 germ and 3 somatic cell subsets. Among the germ cell clusters are 4 undifferentiated spermatogonia (SPG) states, each marked by specific genes. One of the SPG states has characteristics suggesting it is enriched for SSCs. We identified protein markers specific for this state, including cell-surface proteins that we used to enrich for these cells. We mapped the timeline of male germ cell development from PGCs through fetal germ cells to differentiating adult SPG stages. We also defined somatic cell subsets in the human testis and traced their developmental trajectories. Together, our data provides a blueprint for understanding the development of the male germline and supporting somatic cells in humans. The germ cell subset markers we identified are candidates to be used for clinical applications, including SSC therapy for treating infertility.

2018-12-22 | GSE124263 | GEO

Expression data from cultured spermatogonia established from 10 day old mouse testes

Project description:Spermatogonial stem cells (SSCs) undergo self-renewal division to sustain spermatogenesis. Although it is possible to derive germline stem (GS) cell cultures from most of the mouse strains by supplementing GDNF and FGF2, SSCs from a 129 background do not proliferate under the same culture conditions, which suggested that they have distinct self-renewal requirements. We modified previous culture conditions and established long-term culture of SSCs of 129 mice. 129 GS cells reinitiated spermatogenesis and produced offspring following transplantation into the seminiferous tubules of infertile mouse recipients. This dataset show the differences of gene expressions of GS cells between C57BL/6 and 129 mice, which have important implications in understanding requirements of self-renewal mechanisms. In this dataset, we include the expression data obtained from cultured spermatogonia (GS cells) derived from C57BL/6, and 129 mice. Each group contains 2 biological replicates.

2024-06-17 | GSE236119 | GEO

Expression data from spermatogonia enriched from 12 dpc mouse testes

Project description:Spermatogonial stem cells (SSCs) cotinues to produce sperm throughout life by self-renewal division. Although it is possible to expand SSCs from most of the mouse strains in vitro for a long-term period by supplementing GDNF and FGF2, SSCs from a 129 background do not proliferate under the same culture conditions. Since it suggested that they have distinct self-renewal requirements, gene expressions of spermatogonia enriched from testes of DBA/2, C57BL/6, and 129 strains were compared. In this dataset, we include the expression data obtained from spermatogonia enriched from 12dpc testes of DBA/2, C57BL/6, and 129. Enrichment was performed by magnetic activated-cell sorting using anti-Ecadherin (CDH1) antibody (ECCD2). These data are used to obtain 359 genes that are differentially expressed among DBA/2, C57BL/6, and 129.

2024-06-30 | GSE234896 | GEO

Identification of EOMES-expressing spermatogonial stem cells and their regulation by PLZF

Project description:Long-term maintenance of spermatogenesis in mammals is supported by GDNF, an essential growth factor required for spermatogonial stem cell (SSC) self-renewal. Exploiting a transgenic GDNF overexpression model, which expands and normalizes the pool of undifferentiated spermatogonia between Plzf +/+ and Plzf lu/lu mice, we used RNAseq to identify a rare subpopulation of cells that express EOMES, a T-box transcription factor. Lineage tracing, conditional ablation, and busulfan challenge show that these are long-term SSCs that contribute to steady state spermatogenesis as well as regeneration following chemical injury. EOMES+ SSCs have a lower proliferation index than EOMES− GFRA1+ spermatogonia in wild-type but not in Plzf lu/lu mice. This comparison demonstrates that PLZF regulates their proliferative activity and suggests that EOMES+ SSCs are lost through proliferative exhaustion in Plzf lu/lu mice. Single cell RNA sequencing of EOMES+ cells from Plzf +/+ and Plzf lu/lu mice support a hierarchical model of a slow-cycling long-term SSC population supporting more rapid-cycling short-term SSCs.

2019-05-30 | GSE116001 | GEO

Sertoli Cell-Only Syndrome Human Transcriptome Profiling Study

Project description:<p>Sertoli cell-only syndrome is severe form of human male infertility in which most seminiferous tubules appear to lack all spermatogenic cells, including spermatogonial stem cells (SSCs). However, a few small tubule segments of some patients have active spermatogenesis and, thus, functional stem cell niches and SSCs. Normally SSCs replicate, migrate and refill adjacent empty niches, but this does not appear to occur in SCO syndrome. We hypothesized that this failure occurs because most niches are dysfunctional. As Sertoli cells are essential to formation of these niches, we used RNAseq to compare the transcriptomes of human testes with qualitatively normal (complete) spermatogenesis (n=4) with the transcriptomes of human testes with SCO syndrome (n=7). We then focused our analysis on the expression of transcripts that bioinformatic analyses identified as Sertoli cell signature transcripts. Results show that Sertoli cells in SCO testes express abnormally low levels of GDNF, FGF8 and BMP4, all of which are important regulators of mouse SSCs and/or progenitor spermatogonia. Sertoli cells in SCO testes express significantly reduced levels of transcripts for proteins that polarize the Sertoli cell plasma membrane and regulate the trafficking of cell adhesion and gap junction proteins in and out of that plasma membrane.</p>

| phs001777 | dbGaP

Kaiso, a critical regulator of spermatogenesis

Project description:Spermatogenesis is a highly coordinated process by which diploid spermatogonia (2n) differentiate into mature haploid (1n) spermatozoa in the seminiferous epithelium. Here, we show that Kaiso (Zbtb33) is critical to spermatogenesis, via binding to KBEs (Kaiso-binding elements) in the 5’ upstream regulatory regions of Gdnf and Plzf to repress their expression, promoting the differentiation of spermatogonial stem cells (SSCs). In Kaiso-/Y mice, Plzf and Gdnf were derepressed in both SSCs and Sertoli cells, explaining the increased number of SSCs observed in these abnormal mice. We also found that Bcl6, an antiapoptotic spermatocyte survival factor, represses the Crem pathway genes. In particular, Kaiso represses Bcl6 by binding two KBEs in the Bcl6 5’ upstream regulatory region and first intron, thus indirectly derepressing Crem and Crem pathway genes. In Kaiso-/Y mice, Bcl6 is overexpressed, and these mice lack differentiated post-mitotic cells and consequently, are infertile. Interestingly, epigenetic “marks,” histone post-translational modifications such as lysine methylation, within histone H3 (H3K27me3), of the promoters of Kaiso target genes, were found transgenerationally heritable, prior to “wash out” during generations 1-3 in Kaiso-/Y mice, thus impacting Kaiso target gene expression. Consequently, Kaiso, by repressing Plzf, Gdnf, and Bcl6, indirectly upregulates Crem, and thereby plays a critical role not only in SSC differentiation, but also in post-meiotic events during spermatogenesis.

2021-03-05 | GSE111409 | GEO

GDNF-regulated gene expression in cultured mouse spermatogonial stem cells

Project description:GDNF-regulated gene expression was studied in cultures of actively self-renewing spermatogonial stem cells established from 6 day old male mice. GDNF is the essential growth factor regulating mouse spermatogonial stem cell self-renewal. Using a serum-free chemically defined culture system that supports mouse spermatogonial stem cell self-renewal for extended periods of time, GDNF-regulated genes were identified using microarray profiling. Keywords: GDNF withdrawal and time-course replacement

2006-05-11 | GSE4799 | GEO

LncRNA Sequencing Analysis of Mouse spermatogonial stem cells during Glial cell-derived neurotrophic factor（GDNF）treatment

2015-04-15 | GSE66998 | GEO

Dynamics of DNA methylation patterning during in vitro propagation of human spermatogonial stem cells

Project description:Autotransplantation of in vitro proliferated spermatogonial stem cells (SSCs) has been advocated as a way to restore fertility in childhood cancer survivors. In this study we determined the effects of cell culture on DNA methylation patterning in human SSCs, to evaluate the safety of future clinical application of SSC autotransplantation. Bisulfite converted DNA of 34 human spermatogonial stem cell samples, 4 sperm samples and 3 seminoma samples were hybridized to Illumina Infinium 450k Human Methylation Beadchips

2016-02-27 | E-GEOD-72444 | biostudies-arrayexpress

OmicsDI is part of the ELIXIR infrastructure

OmicsDI is an Elixir interoperability service. Learn more ›

Tweets

OmicsDI Databases

PRIDE
PeptideAtlas
MassIVE
JPOST Repository
Physiome Model Repository

EGA
EVA
ENA
LINCS
PAXDB
Cell Collective

MetaboLights
Metabolomics Workbench
MetabolomeExpress
GNPS
BioModels
FAIRDOMHub

ArrayExpress
dbGaP
ExpressionAtlas
GEO
NODE

Information

Databases
Help
API
Contact us
Code on GitHub
Terms of use
Submit Data