Induction of meiosis by embryonic gonadal somatic cells differentiated from pluripotent stem cells

Abstract Background Depletion of oocytes leads to ovarian aging-associated infertility, endocrine disruption and related diseases. Excitingly, unlimited oocytes can be generated by differentiation of primordial germ cell like cells (PGCLCs) from pluripotent stem cells. Nevertheless, development of oocytes and follicles from PGCLCs relies on developmentally matched gonadal somatic cells, only available from E12.5 embryos in mice. It is therefore imperative to achieve an in vitro source of E12.5 gonadal somatic cells. Methods We explored to identify small molecules, which can induce female embryonic stem cells (ESCs) into gonadal somatic cell like cells. Results Using RNA-sequencing, we identified signaling pathways highly upregulated in E12.5_gonadal somatic cells (E12.5_GSCs). Through searching for the activators of these pathways, we identified small-molecule compounds Vitamin C (Vc) and AM580 in combination (V580) for inducing differentiation of female embryonic stem cells (ESCs) into E12.5_GSC-like cells (E12.5_GSCLCs). After V580 treatment for 6 days and sorted by a surface marker CD63, the cell population yielded a transcriptome profile similar to that of E12.5_GSCs, which promoted meiosis progression and folliculogenesis of primordial germ cells. This approach will contribute to the study of germ cell and follicle development and oocyte production and have implications in potentially treating female infertility. Conclusion ESCs can be induced into embryonic gonadal somatic cell like cells by small molecules.

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Testicular somatic cell-like cells derived from embryonic stem cells induce differentiation of epiblasts into germ cells

Communications Biology ◽

10.1038/s42003-021-02322-8 ◽

2021 ◽

Vol 4 (1) ◽

Author(s):

Holly Rore ◽

Nicholas Owen ◽

Raul Eduardo Piña-Aguilar ◽

Kevin Docherty ◽

Ryohei Sekido

Keyword(s):

Stem Cells ◽

Embryonic Stem Cells ◽

Somatic Cell ◽

Germ Cells ◽

Pluripotent Stem Cells ◽

Seminiferous Tubule ◽

Embryonic Stem ◽

In Vitro Reconstitution ◽

Real Challenge

AbstractRegeneration of the testis from pluripotent stem cells is a real challenge, reflecting the complexity of the interaction of germ cells and somatic cells. Here we report the generation of testicular somatic cell-like cells (TesLCs) including Sertoli cell-like cells (SCLCs) from mouse embryonic stem cells (ESCs) in xeno-free culture. We find that Nr5a1/SF1 is critical for interaction between SCLCs and PGCLCs. Intriguingly, co-culture of TesLCs with epiblast-like cells (EpiLCs), rather than PGCLCs, results in self-organised aggregates, or testicular organoids. In the organoid, EpiLCs differentiate into PGCLCs or gonocyte-like cells that are enclosed within a seminiferous tubule-like structure composed of SCLCs. Furthermore, conditioned medium prepared from TesLCs has a robust inducible activity to differentiate EpiLCs into PGCLCs. Our results demonstrate conditions for in vitro reconstitution of a testicular environment from ESCs and provide further insights into the generation of sperm entirely in xeno-free culture.

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Gamete derivation from embryonic stem cells, induced pluripotent stem cells or somatic cell nuclear transfer-derived embryonic stem cells: state of the art

Reproduction Fertility and Development ◽

10.1071/rd14317 ◽

2015 ◽

Vol 27 (1) ◽

pp. 89 ◽

Cited By ~ 11

Author(s):

Charles A. Easley ◽

Calvin R. Simerly ◽

Gerald Schatten

Keyword(s):

Stem Cells ◽

Embryonic Stem Cells ◽

Somatic Cell ◽

Germ Cells ◽

Pluripotent Stem Cells ◽

Primordial Germ Cells ◽

Embryonic Stem ◽

Induced Pluripotent ◽

Normal Offspring

Generating gametes from pluripotent stem cells (PSCs) has many scientific justifications and several biomedical rationales. Here, we consider several strategies for deriving gametes from PSCs from mice and primates (human and non-human) and their anticipated strengths, challenges and limitations. Although the ‘Weismann barrier’, which separates the mortal somatic cell lineages from the potentially immortal germline, has long existed, breakthroughs first in mice and now in humans are artificially creating germ cells from somatic cells. Spermatozoa with full reproductive viability establishing multiple generations of seemingly normal offspring have been reported in mice and, in humans, haploid spermatids with correct parent-of-origin imprints have been obtained. Similar progress with making oocytes has been published using mouse PSCs differentiated in vitro into primordial germ cells, which are then cultured after xenografting reconstructed artificial ovaries. Progress in making human oocytes artificially is proving challenging. The usefulness of these artificial gametes, from assessing environmental exposure toxicity to optimising medical treatments to prevent negative off-target effects on fertility, may prove invaluable, as may basic discoveries on the fundamental mechanisms of gametogenesis.

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Induction of the germ cell fate from pluripotent stem cells in cynomolgus monkeys†

Biology of Reproduction ◽

10.1093/biolre/ioz205 ◽

2019 ◽

Vol 102 (3) ◽

pp. 620-638 ◽

Cited By ~ 5

Author(s):

Yoshitake Sakai ◽

Tomonori Nakamura ◽

Ikuhiro Okamoto ◽

Sayuri Gyobu-Motani ◽

Hiroshi Ohta ◽

...

Keyword(s):

Stem Cells ◽

Germ Cell ◽

Cell Fate ◽

Pluripotent Stem Cells ◽

Primordial Germ Cell ◽

Embryonic Stem ◽

Cell Development ◽

Cynomolgus Monkeys ◽

Germ Cell Development

Abstract In vitro reconstitution of germ-cell development from pluripotent stem cells (PSCs) has created key opportunities to explore the fundamental mechanisms underlying germ-cell development, particularly in mice and humans. Importantly, such investigations have clarified critical species differences in the mechanisms regulating mouse and human germ-cell development, highlighting the necessity of establishing an in vitro germ-cell development system in other mammals, such as non-human primates. Here, we show that multiple lines of embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) in cynomolgus monkeys (Macaca fascicularis; cy) can be maintained stably in an undifferentiated state under a defined condition with an inhibitor for WNT signaling, and such PSCs are induced efficiently into primordial germ cell-like cells (PGCLCs) bearing a transcriptome similar to early cyPGCs. Interestingly, the induction kinetics of cyPGCLCs from cyPSCs is faster than that of human (h) PGCLCs from hPSCs, and while the transcriptome dynamics during cyPGCLC induction is relatively similar to that during hPGCLC induction, it is substantially divergent from that during mouse (m) PGCLC induction. Our findings delineate common as well as species-specific traits for PGC specification, creating a foundation for parallel investigations into the mechanism for germ-cell development in mice, monkeys, and humans.

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A Germ Cell-specific Gene, Prmt5, Works in Somatic Cell Reprogramming

Journal of Biological Chemistry ◽

10.1074/jbc.m110.216390 ◽

2011 ◽

Vol 286 (12) ◽

pp. 10641-10648 ◽

Cited By ~ 48

Author(s):

Go Nagamatsu ◽

Takeo Kosaka ◽

Miyuri Kawasumi ◽

Taisuke Kinoshita ◽

Keiyo Takubo ◽

...

Keyword(s):

Stem Cells ◽

Germ Cell ◽

Somatic Cell ◽

Germ Cells ◽

Pluripotent Stem Cells ◽

Somatic Cells ◽

Embryonic Stem ◽

Cell Reprogramming ◽

Specific Gene ◽

Somatic Cell Reprogramming

Germ cells possess the unique ability to acquire totipotency during development in vivo as well as give rise to pluripotent stem cells under the appropriate conditions in vitro. Recent studies in which somatic cells were experimentally converted into pluripotent stem cells revealed that genes expressed in primordial germ cells (PGCs), such as Oct3/4, Sox2, and Lin28, are involved in this reprogramming. These findings suggest that PGCs may be useful for identifying factors that successfully and efficiently reprogram somatic cells into toti- and/or pluripotent stem cells. Here, we show that Blimp-1, Prdm14, and Prmt5, each of which is crucial for PGC development, have the potential to reprogram somatic cells into pluripotent stem cells. Among them, Prmt5 exhibited remarkable reprogramming of mouse embryonic fibroblasts into which Prmt5, Klf4, and Oct3/4 were introduced. The resulting cells exhibited pluripotent gene expression, teratoma formation, and germline transmission in chimeric mice, all of which were indistinguishable from those induced with embryonic stem cells. These data indicate that some of the factors that play essential roles in germ cell development are also active in somatic cell reprogramming.

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In Vitro Differentiation of T Cells: From Human Embryonic Stem Cells and Induced Pluripotent Stem Cells

Methods in Molecular Biology - In Vitro Differentiation of T-Cells ◽

10.1007/978-1-4939-9728-2_6 ◽

2019 ◽

pp. 59-70 ◽

Cited By ~ 1

Author(s):

Shoichi Iriguchi ◽

Shin Kaneko

Keyword(s):

Stem Cells ◽

T Cells ◽

Embryonic Stem Cells ◽

Induced Pluripotent Stem Cells ◽

Human Embryonic Stem Cells ◽

Pluripotent Stem Cells ◽

Embryonic Stem ◽

In Vitro Differentiation ◽

Induced Pluripotent

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SET domain-containing protein 5 is required for expression of primordial germ cell specification-associated genes in murine embryonic stem cells

Cell Biochemistry and Function ◽

10.1002/cbf.3269 ◽

2017 ◽

Vol 35 (5) ◽

pp. 247-253 ◽

Cited By ~ 4

Author(s):

Seung Eun Yu ◽

Min Seong Kim ◽

Su Hyung Park ◽

Byong Chul Yoo ◽

Kyung Hee Kim ◽

...

Keyword(s):

Stem Cells ◽

Embryonic Stem Cells ◽

Germ Cell ◽

Primordial Germ Cell ◽

Embryonic Stem ◽

Set Domain ◽

Cell Specification ◽

Murine Embryonic Stem Cells ◽

Germ Cell Specification

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Gene Expression and Epigenetic Signatures of Germ Cell-Derived Pluripotent Stem Cells and Embryonic Stem Cells

Stem Cells and Cancer Stem Cells, Volume 6 ◽

10.1007/978-94-007-2993-3_3 ◽

2012 ◽

pp. 21-34

Author(s):

Jessica Nolte ◽

D. V. Krishna Pantakani ◽

Hassan Dihazi ◽

Ulrich Zechner

Keyword(s):

Gene Expression ◽

Stem Cells ◽

Embryonic Stem Cells ◽

Germ Cell ◽

Pluripotent Stem Cells ◽

Embryonic Stem ◽

Epigenetic Signatures

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Cyclosporin A promotes invasion and migration of extravillous trophoblast cells derived from human induced pluripotent stem cells and human embryonic stem cells

10.21203/rs.3.rs-25667/v1 ◽

2020 ◽

Author(s):

Jiaxing Wang ◽

Ping Long ◽

Shengnan Tian ◽

Weihua Zu ◽

Jing Liu ◽

...

Keyword(s):

Stem Cells ◽

Embryonic Stem Cells ◽

Human Embryonic Stem Cells ◽

Pluripotent Stem Cells ◽

Embryonic Stem ◽

Extravillous Trophoblast ◽

Invasion And Migration ◽

And Migration ◽

Induced Pluripotent

Abstract Background Extravillous trophoblast (EVT) cells play an essential role in the maternal-fetal interaction. Although abnormal development and function of EVT cells, including impaired migration and invasion capability, are believed to be etiologically linked to severe pregnancy disorders including pre-eclampsia (PE), the associated molecular mechanisms are not clear ascribed to the lack of an appropriate cell model in vitro. Cyclosporine A (CsA) is a macrolide immunosuppressant and is also used in clinic to improve pregnancy outcomes. However, whether CsA has any effects on the function of EVT cells has not been well investigated. Methods In this study, we induced differentiation of human induced pluripotent stem cells (hiPSCs) and human embryonic stem cells (hESCs) into EVT cells (hiPSC-EVT and hESC-EVT cells, respectively) by Y27632, NRG1, A83-01 and matrigel, and collected these derived EVT cells by flow cytometry for sorting cells positive for double HLA-G and KRT7, which are EVT markers. We then investigated the effects of CsA on the invasion and migration of these derived EVT cells. Results We found that the hiPSC-EVT and hESC-EVT cells expressed high levels of the EVT markers such as KRT7, ITGA5 and HLA-G but low levels of OCT4, a stem cell marker, and that CsA significantly promoted the invasion and migration of hiPSC-EVT and hESC-EVT cells. Conclusions We successfully generated hiPSC/hESC-derived human EVT cells, which may be applicable for investigating the remodeling process of spiral arteries remodeling and the possible mechanisms of EVT-related diseases in vitro. Furthermore, our findings provide direct evidence that CsA regulates the function of EVT cells and molecular basis by which CsA may be used to treat pregnancy complications in clinic associated with deficient EVT function.

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Human Cell Modeling for Cardiovascular Diseases

International Journal of Molecular Sciences ◽

10.3390/ijms21176388 ◽

2020 ◽

Vol 21 (17) ◽

pp. 6388

Author(s):

Melania Lippi ◽

Ilaria Stadiotti ◽

Giulio Pompilio ◽

Elena Sommariva

Keyword(s):

Stem Cells ◽

Embryonic Stem Cells ◽

Cardiovascular Diseases ◽

Human Cell ◽

Pluripotent Stem Cells ◽

Disease Modeling ◽

Embryonic Stem ◽

Cell Models ◽

Advantages And Disadvantages

The availability of appropriate and reliable in vitro cell models recapitulating human cardiovascular diseases has been the aim of numerous researchers, in order to retrace pathologic phenotypes, elucidate molecular mechanisms, and discover therapies using simple and reproducible techniques. In the past years, several human cell types have been utilized for these goals, including heterologous systems, cardiovascular and non-cardiovascular primary cells, and embryonic stem cells. The introduction of induced pluripotent stem cells and their differentiation potential brought new prospects for large-scale cardiovascular experiments, bypassing ethical concerns of embryonic stem cells and providing an advanced tool for disease modeling, diagnosis, and therapy. Each model has its advantages and disadvantages in terms of accessibility, maintenance, throughput, physiological relevance, recapitulation of the disease. A higher level of complexity in diseases modeling has been achieved with multicellular co-cultures. Furthermore, the important progresses reached by bioengineering during the last years, together with the opportunities given by pluripotent stem cells, have allowed the generation of increasingly advanced in vitro three-dimensional tissue-like constructs mimicking in vivo physiology. This review provides an overview of the main cell models used in cardiovascular research, highlighting the pros and cons of each, and describing examples of practical applications in disease modeling.

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