Haploid mouse germ cell precursors from embryonic stem cells reveal Xist activation from a single X chromosome

Aizawa, Eishi; Kaufmann, Corinne; Sting, Sarah; Boigner, Sarah; Freimann, Remo; Minin, Giulio Di; Wutz, Anton

doi:10.1016/j.stemcr.2021.11.006

Cited by 3 publications

(5 citation statements)

References 28 publications

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“…This is consistent with the results of a previous study showing that Xist is not expressed in haploid ESCs. However, silencing Xist was not found to improve the maintenance of a haploid genome 45 . Taken together, there is no need for X chromosome inactivation in haESCs, likely because there is only one X chromosome in these cells.…”

Section: Resultsmentioning

confidence: 91%

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Transcriptome profiles and chromatin states in mouse androgenetic haploid embryonic stem cells

Zheng

Wang

et al. 2023

Cell Proliferation

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Haploid embryonic stem cells (haESCs) are derived from the inner cell mass of the haploid blastocyst, containing only one set of chromosomes. Extensive and accurate chromatin remodelling occurs during haESC derivation, but the intrinsic transcriptome profiles and chromatin structure of haESCs have not been fully explored. We profiled the transcriptomes, nucleosome positioning, and key histone modifications of four mouse haESC lines, and compared these profiles with those of other closely‐related stem cell lines, MII oocytes, round spermatids, sperm, and mouse embryonic fibroblasts. haESCs had transcriptome profiles closer to those of naïve pluripotent stem cells. Consistent with the one X chromosome in haESCs, Xist was repressed, indicating no X chromosome inactivation. haESCs and ESCs shared a similar global chromatin structure. However, a nucleosome depletion region was identified in 2056 promoters in ESCs, which was absent in haESCs. Furthermore, three characteristic spatial relationships were formed between transcription factor motifs and nucleosomes in both haESCs and ESCs, specifically in the linker region, on the nucleosome central surface, and nucleosome borders. Furthermore, the chromatin state of 4259 enhancers was off in haESCs but active in ESCs. Functional annotation of these enhancers revealed enrichment in regulation of the cell cycle, a predominantly reported mechanism of haESC self‐diploidization. Notably, the transcriptome profiles and chromatin structure of haESCs were highly preserved during passaging but different from those of differentiated cell types.

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Section: Resultsmentioning

confidence: 91%

“…Xist was not found to improve the maintenance of a haploid genome. 45 Taken together, there is no need for X chromosome inactivation in haESCs, likely because there is only one X chromosome in these cells. Furthermore, X chromosome activation supplies an epigenetic signature of naïve pluripotency.…”

Section: Resultsmentioning

confidence: 99%

Transcriptome profiles and chromatin states in mouse androgenetic haploid embryonic stem cells

Zheng

Wang

et al. 2023

Cell Proliferation

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“…The slightly lower number of beating areas per SP-derived EBs compared to the HD technique can be tolerated due to the much higher throughput and lower workload of the SP technique. The advantages of these plates have also been used by other research groups, for example in ESC and MSC research (66,67), but also in organ-specific tissue engineering (68)(69)(70).…”

Section: Discussionmentioning

confidence: 99%

Heart rhythm in vitro: measuring stem cell-derived pacemaker cells on microelectrode arrays

Kussauer,

Dilk,

Elleisy

et al. 2024

Front. Cardiovasc. Med.

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BackgroundCardiac arrhythmias have markedly increased in recent decades, highlighting the urgent need for appropriate test systems to evaluate the efficacy and safety of new pharmaceuticals and the potential side effects of established drugs.MethodsThe Microelectrode Array (MEA) system may be a suitable option, as it provides both real-time and non-invasive monitoring of cellular networks of spontaneously active cells. However, there is currently no commercially available cell source to apply this technology in the context of the cardiac conduction system (CCS). In response to this problem, our group has previously developed a protocol for the generation of pure functional cardiac pacemaker cells from mouse embryonic stem cells (ESCs). In addition, we compared the hanging drop method, which was previously utilized, with spherical plate-derived embryoid bodies (EBs) and the pacemaker cells that are differentiated from these.ResultsWe described the application of these pacemaker cells on the MEA platform, which required a number of crucial optimization steps in terms of coating, dissociation, and cell density. As a result, we were able to generate a monolayer of pure pacemaker cells on an MEA surface that is viable and electromechanically active for weeks. Furthermore, we introduced spherical plates as a convenient and scalable method to be applied for the production of induced sinoatrial bodies.ConclusionWe provide a tool to transfer modeling and analysis of cardiac rhythm diseases to the cell culture dish. Our system allows answering CCS-related queries within a cellular network, both under baseline conditions and post-drug exposure in a reliable and affordable manner. Ultimately, our approach may provide valuable guidance not only for cardiac pacemaker cells but also for the generation of an MEA test platform using other sensitive non-proliferating cell types.

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“…Recently, a research team used mouse haploid embryonic stem cells to study the activation process of Xist [ 43 ]. Li et al [ 44 ] successfully constructed mouse rat allogeneic hybrid diploid embryonic stem cells (AdESCs) by fusion of mouse and rat haESCs.…”

Section: Haploid Applicationmentioning

confidence: 99%

“…They also systematically analyzed a large number of RNA-seq data and found 146 unknown mouse genes in heterodiploid somatic cells, which may be related to X chromosome inactivation escape. The other research team [ 43 ] obtained haploid primordial germ cell-like cells (PGCLCs) from haploid mouse embryonic stem cells (ESCs). They find that haploid cells show predisposition for PGCLCs, whereas a large fraction of somatic cells becomes diploid.…”

Section: Haploid Applicationmentioning

confidence: 99%

Development and application of haploid embryonic stem cells

Wang,

Ma,

Guo

2024

Stem Cell Res Ther

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Haploid cells are a kind of cells with only one set of chromosomes. Compared with traditional diploid cells, haploid cells have unique advantages in gene screening and drug-targeted therapy, due to their phenotype being equal to the genotype. Embryonic stem cells are a kind of cells with strong differentiation potential that can differentiate into various types of cells under specific conditions in vitro. Therefore, haploid embryonic stem cells have the characteristics of both haploid cells and embryonic stem cells, which makes them have significant advantages in many aspects, such as reproductive developmental mechanism research, genetic screening, and drug-targeted therapy. Consequently, establishing haploid embryonic stem cell lines is of great significance. This paper reviews the progress of haploid embryonic stem cell research and briefly discusses the applications of haploid embryonic stem cells.

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Haploid mouse germ cell precursors from embryonic stem cells reveal Xist activation from a single X chromosome

Cited by 3 publications

References 28 publications

Transcriptome profiles and chromatin states in mouse androgenetic haploid embryonic stem cells

Transcriptome profiles and chromatin states in mouse androgenetic haploid embryonic stem cells

Heart rhythm in vitro: measuring stem cell-derived pacemaker cells on microelectrode arrays

Development and application of haploid embryonic stem cells

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