Single-cell gene expression analysis reveals diversity among human spermatogonia

Neuhaus, Nina; Yoon, Jong Ho; Terwort, Nicole; Kliesch, Sabine; Seggewiß, Jochen; Huge, Andreas; Voss, Randal; Schlatt, Stefan; Grindberg, Rashel V.; Schöler, Hans R.

doi:10.1093/molehr/gaw079

Cited by 32 publications

(34 citation statements)

References 40 publications

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“…In contrast to UTF1, UCH-L1 is expressed in all Ap-d spermatogonia and, similarly to UTF1, UCH-L1 expression is lost in B spermatogonia. Recently, a single cell RNA-seq experiment was carried out on human spermatogonia (Neuhaus et al, 2017). Heterogeneous expression profiles were found but the results did not yet lead to a better insight into spermatogonial behavior in humans.…”

Section: Discussionmentioning

confidence: 99%

Spermatogonial kinetics in humans

et al. 2017

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The human spermatogonial compartment is essential for daily production of millions of sperm. Despite this crucial role, the molecular signature, kinetic behavior and regulation of human spermatogonia are poorly understood. Using human testis biopsies with normal spermatogenesis and by studying marker protein expression, we have identified for the first time different subpopulations of spermatogonia. MAGE-A4 marks all spermatogonia, KIT marks all B spermatogonia and UCLH1 all Apale-dark (Ap-d) spermatogonia. We suggest that at the start of the spermatogenic lineage there are Ap-d spermatogonia that are GFRA1, likely including the spermatogonial stem cells. Next, UTF1 becomes expressed, cells become quiescent and GFRA1 expression decreases. Finally, GFRA1 expression is lost and subsequently cells differentiate into B spermatogonia, losing UTF1 and acquiring KIT expression. Strikingly, most human Ap-d spermatogonia are out of the cell cycle and even differentiating type B spermatogonial proliferation is restricted. A novel scheme for human spermatogonial development is proposed that will facilitate further research in this field, the understanding of cases of infertility and the development of methods to increase sperm output.

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Section: Discussionmentioning

confidence: 99%

Spermatogonial kinetics in humans

et al. 2017

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“…There is some evidence of the role of epigenetic factors in the regulation of sperm development and quality [73] and the potential for transgenerational inheritance of epigenetic germ line mutations. As such, we strongly feel that more studies are required that focus on transcriptomic and epigenetic measurements in human germ cell cultures, for example through the use of novel emerging single-cell sequencing approaches [74][75][76], to gain insight in the events that occur during in vitro culture of human testicular cells. and the X chromosome as aggregates of each sample group (see Methods).…”

Section: Plos Onementioning

confidence: 99%

Comparing genome-scale DNA methylation and CNV marks between adult human cultured ITGA6+ testicular cells and seminomas to assess in vitro genomic stability

et al. 2020

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Autologous transplantation of spermatogonial stem cells is a promising new avenue to restore fertility in infertile recipients. Expansion of the initial spermatogonial stem cell pool through cell culturing is a necessary step to obtain enough cells for effective repopulation of the testis after transplantation. Since in vitro propagation can lead to (epi-)genetic mutations and possibly malignant transformation of the starting cell population, we set out to investigate genome-wide DNA methylation status in uncultured and cultured primary testicular ITGA6+ sorted cells and compare them with germ cell tumor samples of the seminoma subtype. Seminomas displayed a severely global hypomethylated profile, including loss of genomic imprinting, which we did not detect in cultured primary testicular ITGA6+ cells. Differential methylation analysis revealed altered regulation of gamete formation and meiotic processes in cultured primary testicular ITGA6+ cells but not in seminomas. The pivotal POU5F1 marker was hypomethylated in seminomas but not in uncultured or cultured primary testicular ITGA6+ cells, which is reflected in the POU5F1 mRNA expression levels. Lastly, seminomas displayed a number of characteristic copy number variations that were not detectable in primary testicular ITGA6+ cells, either before or after culture. Together, the data show a distinct DNA methylation patterns in cultured primary testicular ITGA6+ cells that does not resemble the pattern found in seminomas, but also highlight the need for more sensitive methods to fully exclude the presence of malignant cells after culture and to further study the epigenetic events that take place during in vitro culture. PLOS ONE PLOS ONE | https://doi.org/10.1371/journal.pone.0230253 March 16, 2020 1 / 18 OPEN ACCESS Citation: Struijk RB, Dorssers LCJ, Henneman P, Rijlaarsdam MA, Venema A, Jongejan A, et al. (2020) Comparing genome-scale DNA methylation and CNV marks between adult human cultured ITGA6+ testicular cells and seminomas to assess in vitro genomic stability. PLoS ONE 15(3): e0230253. https://doi.org/10.

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“…Over the past few years, molecular tracing experiments coupled with single-cell mRNA sequencing and transplantation assays uncovered the fact that the mammalian A single cell population is heterogeneous (Grisanti et al 2009, Chan et al 2014, Hermann et al 2015, Mutoji et al 2016, Guo et al 2017, Neuhaus et al 2017, Green et al 2018, and that subsets of A single cells have different capacity for self-renewal (Aloisio et al 2014, Helsel et al 2017. According to these data, a new model is now emerging whereby in mice a subpopulation of A single cells, marked by high expression of both the transcription factor ID4 and the membrane receptor GFRA1, a co-receptor for GDNF, is the purest functional SSC population described to date in immature and adult mice (Sun et al 2015, Lord & Oatley 2017, Hermann et al 2018.…”

Section: Introductionmentioning

confidence: 99%

Regulation of GDNF expression in Sertoli cells

2019

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Sertoli cells regulate male germ cell proliferation and differentiation and are a critical component of the spermatogonial stem cell (SSC) niche, where homeostasis is maintained by the interplay of several signaling pathways and growth factors. These factors are secreted by Sertoli cells located within the seminiferous epithelium, and by interstitial cells residing between the seminiferous tubules. Sertoli cells and peritubular myoid cells produce glial cell line-derived neurotrophic factor (GDNF), which binds to the RET/GFRA1 receptor complex at the surface of undifferentiated spermatogonia. GDNF is known for its ability to drive SSC self-renewal and proliferation of their direct cell progeny. Even though the effects of GDNF are well studied, our understanding of the regulation its expression is still limited. The purpose of this review is to discuss how GDNF expression in Sertoli cells is modulated within the niche, and how these mechanisms impact germ cell homeostasis.

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Single-cell gene expression analysis reveals diversity among human spermatogonia

Abstract: This work was supported by the Max Planck Society and the Deutsche Forschungsgemeinschaft DFG-Research Unit FOR 1041 Germ Cell Potential (grant numbers SCHO 340/7-1, SCHL394/11-2). The authors declare that there is no conflict of interest.

Cited by 32 publications

References 40 publications

Spermatogonial kinetics in humans

Spermatogonial kinetics in humans

Comparing genome-scale DNA methylation and CNV marks between adult human cultured ITGA6+ testicular cells and seminomas to assess in vitro genomic stability

Regulation of GDNF expression in Sertoli cells

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