In Vitro Meiosis of Male Germline Stem Cells

Lei, Qijing; Lai, Xinmin; Eliveld, Jitske; Lopes, Susana M. Chuva de Sousa; Pelt, Ans M.M. van; Hamer, Geert

doi:10.1016/j.stemcr.2020.10.006

Cited by 20 publications

(43 citation statements)

References 65 publications

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“…In accordance with the very inefficient formation of meiotic crossovers, these M-phase cells only displayed pairs of sister chromatids (univalents) with CREST-stained centromeres located at the ends of the paired sister chromatids (Figure 4A). In addition, 1-2 flower-shaped cells per microscope slide, identified as a type of premature M-phase cells in our recent study (17), could still be observed between day 8 to day 15 in this current culture system (Figure 4B). In all, we conclude that, despite the completion of synapsis and XY body formation, meiotic crossover formation was still not completed in vitro, which prevents formation of bivalents (pairs of homologous chromosomes) during the first meiotic M-phase.…”

Section: Meiotic Recombination Is Not Completed In Vitrosupporting

confidence: 57%

“…In Vitro Meiosis of mGSCs SK49 cells, inactivated by mitomycin (10µg/mL, M7949, Sigma), were grown on 12-well plates pre-coated with laminin (20 µg/ mL, L2020, Sigma) to a density of 1 x10 5 per well. Then mGSCs were seeded on these Sertoli cells to maintain mGSCs proliferation for two days at 37°C using medium I, composed of as described previously (17,29). To induce meiosis, the cells were cultured at 34°C.…”

Section: Male Germline Stem Cells and Sertoli Cell Line Culturementioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

“…Several studies have described complete spermatogenesis in vitro using various cell culture strategies and starting cell types, such as pluripotent stem cells (PSCs) (2,(18)(19)(20)(21), or spermatogonial stem cells (SSCs) (17,(22)(23)(24). Of these studies only few characterized whether key meiotic events took place in vitro (2,17). When mouse embryonic stem cells (ESCs), grown on a cell suspension of male neonatal gonad, were induce to complete in vitro meiosis, no meiotic problems were reported (2).…”

Section: Introductionmentioning

confidence: 99%

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Meiotic Chromosome Synapsis and XY-Body Formation In Vitro

et al. 2021

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To achieve spermatogenesis in vitro, one of the most challenging processes to mimic is meiosis. Meiotic problems, like incomplete synapsis of the homologous chromosomes, or impaired homologous recombination, can cause failure of crossover formation and subsequent chromosome nondisjunction, eventually leading to aneuploid sperm. These meiotic events are therefore strictly monitored by meiotic checkpoints that initiate apoptosis of aberrant spermatocytes and lead to spermatogenic arrest. However, we recently found that, in vitro derived meiotic cells proceeded to the first meiotic division (MI) stage, despite displaying incomplete chromosome synapsis, no discernible XY-body and lack of crossover formation. We therefore optimized our in vitro culture system of meiosis from male germline stem cells (mGSCs) in order to achieve full chromosome synapsis, XY-body formation and meiotic crossovers. In comparison to previous culture system, the in vitro-generated spermatocytes were transferred after meiotic initiation to a second culture dish. This dish already contained a freshly plated monolayer of proliferatively inactivated immortalized Sertoli cells supporting undifferentiated mGSCs. In this way we aimed to simulate the multiple layers of germ cell types that support spermatogenesis in vivo in the testis. We found that in this optimized culture system, although independent of the undifferentiated mGSCs, meiotic chromosome synapsis was complete and XY body appeared normal. However, meiotic recombination still occurred insufficiently and only few meiotic crossovers were formed, leading to MI-spermatocytes displaying univalent chromosomes (paired sister chromatids). Therefore, considering that meiotic checkpoints are not necessarily fully functional in vitro, meiotic crossover formation should be closely monitored when mimicking gametogenesis in vitro to prevent generation of aneuploid gametes.

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Section: Meiotic Recombination Is Not Completed In Vitrosupporting

confidence: 57%

Section: Male Germline Stem Cells and Sertoli Cell Line Culturementioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Meiotic Chromosome Synapsis and XY-Body Formation In Vitro

et al. 2021

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“…In future, development of single-cell Hi-C technology would help to unravel the finer 3D chromatin structural difference between SSCs and progenitors, enabling more comprehensive insights into the higher-order chromatin dynamics during male germline development, and with functional perturbation analyses, the roles of 3D genome conformation in transcriptional activity could be validated. Overall, the present study adds to the growing body of knowledge about chromatin configuration related to male fertility, and may potentially contribute to treatment of male infertility by SSC therapy, i.e., SSC auto-transplantation (Mulder et al, 2016) or in vitro differentiation into sperm (Lei et al, 2020).…”

Section: Discussionmentioning

confidence: 83%

Unraveling three-dimensional chromatin structural dynamics during spermatogonial differentiation

Zheng

Zhang

Jin

et al. 2021

Preprint

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Spermatogonial stem cells (SSCs) are able to undergo self-renewal and differentiation. Unlike the self-renewal that replenishes the SSC and progenitor pool, the differentiation is an irreversible process committed to meiosis. While the preparations for meiotic events in differentiating spermatogonia (Di-SG) are likely to be accompanied by alterations in chromatin structure, the three-dimensional (3D) chromatin architectural difference between SSCs and Di-SG, and the higher-order chromatin dynamics during spermatogonial differentiation, have not been systematically investigated. Here, we performed in situ high throughput chromosome conformation capture (Hi-C), RNA-sequencing (RNA-seq) and chromatin immunoprecipitation-sequencing (ChIP-seq) analyses on porcine undifferentiated spermatogonia (Un-SG, which consist of SSCs and progenitors) and Di-SG. By integrating and analyzing these data, we identified that Di-SG exhibited increased disorder but weakened compartmentalization and topologically associating domains (TADs) in comparison with Un-SG, suggesting that diminished higher-order chromatin architecture in meiotic cells, as shown by recent reports, is preprogramed in Di-SG. Our data also revealed that A/B compartments and TADs were related to dynamic gene expression during spermatogonial differentiation. We further unraveled the contribution of promoter-enhancer interactions (PEIs) to pre-meiotic transcriptional regulation, which has not been accomplished in previous studies due to limited cell input and resolution. Together, our study uncovered the 3D chromatin structure of SSCs/progenitors and Di-SG, as well as the interplay between higher-order chromatin architecture and dynamic gene expression during spermatogonial differentiation, providing novel insights into the mechanisms for SSC self-renewal and differentiation and having implications for diagnosis and treatment of male sub-/infertility.

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How germline genes promote malignancy in cancer cells

et al. 2022

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Cancers often express hundreds of genes otherwise specific to germ cells, the germline/cancer (GC) genes. Here, we present and discuss the hypothesis that activation of a "germline program" promotes cancer cell malignancy. We do so by proposing four hallmark processes of the germline: meiosis, epigenetic plasticity, migration, and metabolic plasticity. Together, these hallmarks enable replicative immortality of germ cells as well as cancer cells. Especially meiotic genes are frequently expressed in cancer, implying that genes unique to meiosis may play a role in oncogenesis. Because GC genes are not expressed in healthy somatic tissues, they form an appealing source of specific treatment targets with limited side effects besides infertility. Although it is still unclear why germ cell specific genes are so abundantly expressed in cancer, from our hypothesis it follows that the germline's reproductive program is intrinsic to cancer development.

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In Vitro Meiosis of Male Germline Stem Cells

Cited by 20 publications

References 65 publications

Meiotic Chromosome Synapsis and XY-Body Formation In Vitro

Meiotic Chromosome Synapsis and XY-Body Formation In Vitro

Unraveling three-dimensional chromatin structural dynamics during spermatogonial differentiation

How germline genes promote malignancy in cancer cells

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