In vitro reconstitution of the whole male germ-cell development from mouse pluripotent stem cells

Reconstitution in culture of biological processes, such as differentiation and organization, is a key challenge in regenerative medicine, and one in which stem cell technology plays a central role. Pluripotent stem cells and spermatogonial stem cells are useful materials for reconstitution of germ cell development in vitro, as they are capable of differentiating into gametes. Reconstitution of germ cell development, termed in vitro gametogenesis, will provide an experimental platform for a better understanding of germ cell development, as well as an alternative source of gametes for reproduction, with the potential to cure infertility. Since germ cells are the cells for ‘the next generation’, both the culture system and its products must be carefully evaluated. In this issue, we summarize the progress in in vitro gametogenesis, most of which has been made using mouse models, as well as the future challenges in this field.

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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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In Vitro Modeling of Human Germ Cell Development Using Pluripotent Stem Cells

Stem Cell Reports ◽

10.1016/j.stemcr.2018.01.001 ◽

2018 ◽

Vol 10 (2) ◽

pp. 509-523 ◽

Cited By ~ 19

Author(s):

Yuncheng Zhao ◽

Shicheng Ye ◽

Dongli Liang ◽

Pengxiang Wang ◽

Jing Fu ◽

...

Keyword(s):

Stem Cells ◽

Germ Cell ◽

Pluripotent Stem Cells ◽

Cell Development ◽

In Vitro Modeling ◽

Germ Cell Development

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Mammalian germ cell development: from mechanism to in vitro reconstitution

Stem Cell Reports ◽

10.1016/j.stemcr.2021.01.008 ◽

2021 ◽

Author(s):

Mitinori Saitou

Keyword(s):

Germ Cell ◽

Cell Development ◽

Germ Cell Development ◽

In Vitro Reconstitution

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Artificially produced gametes in mice, humans and other species

Reproduction Fertility and Development ◽

10.1071/rd20265 ◽

2021 ◽

Vol 33 (2) ◽

pp. 91

Author(s):

Katsuhiko Hayashi ◽

Cesare Galli ◽

Sebastian Diecke ◽

Thomas B. Hildebrandt

Keyword(s):

Stem Cells ◽

Germ Cell ◽

Pluripotent Stem Cells ◽

Mammalian Species ◽

Cell Development ◽

Current Status ◽

Alternative Source ◽

Germ Cell Development ◽

Further Development

The production of gametes from pluripotent stem cells in culture, also known as invitro gametogenesis, will make an important contribution to reproductive biology and regenerative medicine, both as a unique tool for understanding germ cell development and as an alternative source of gametes for reproduction. Invitro gametogenesis was developed using mouse pluripotent stem cells but is increasingly being applied in other mammalian species, including humans. In principle, the entire process of germ cell development is nearly reconstitutable in culture using mouse pluripotent stem cells, although the fidelity of differentiation processes and the quality of resultant gametes remain to be refined. The methodology in the mouse system is only partially applicable to other species, and thus it must be optimised for each species. In this review, we update the current status of invitro gametogenesis in mice, humans and other animals, and discuss challenges for further development of this technology.

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006. HUMAN GERM CELL FORMATION AND DIFFERENTIATION FROM PLURIPOTENT STEM CELLS

Reproduction Fertility and Development ◽

10.1071/srb10abs006 ◽

2010 ◽

Vol 22 (9) ◽

pp. 5

Author(s):

R. A. ReijoPiera

Keyword(s):

Stem Cells ◽

Germ Cell ◽

Germ Cells ◽

Pluripotent Stem Cells ◽

Rna Binding ◽

Cell Formation ◽

Cell Development ◽

Model Organisms ◽

Germ Cell Development ◽

Germ Cell Formation

Human embryo development begins with the fusion of egg and sperm, followed by reprogramming of the DNA, a series of cell divisions and activation of the embryo’s genome. As development continues, the germ cells (egg and sperm) must be set aside from other cell types. A major cause of infertility in men and women is quantitative and qualitative defects in human germ cell (oocyte and sperm) development. Yet, it has been difficult to study human germ cell development, especially features that are unique relative to model organisms. We have developed a system to differentiate human embryonic stem cells (hESCs) and induced pluripotent stem cells (iPSCs) to germ cells and to quantitate and isolate primordial germ cells (PGCs) derived from both XX- and XY-bearing hESCs and iPSCs. This allowed silencing and overexpression of genes that encode germ cell-specific cytoplasmic RNA-binding proteins (not transcription factors) and resulted in the modulation of human male and female germ cell formation and developmental progression. We observed that human DAZL (Deleted in AZoospermia-Like) functions in female and male PGC formation and maintenance, whereas closely-related family members, BOULE and DAZ, promote entry into meiosis and development of haploid gametes with sperm-specific methylation patterns at imprinted loci in the male. We also conducted critical proof-of-concept studies in mice that showed that phenotypes observed in germ cell development in vitro from wildtype, heterozygous, and Dazl–/– mutation-carrying mouse ESCs (mESCs) mirrored the phenotypes that were observed in vivo. Furthermore, transplantation of XX mESC-derived oocytes resulted in recruitment of somatic cells to form follicles. These studies comprised the first direct experimental analysis of the genetics of human germ cell development and set the stage for extensive exploration of complex genetic variants linked to infertility. Results are significant to the generation of gametes for developmental genetic studies and potential clinical applications.

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In vitro reconstitution of germ cell development†

Biology of Reproduction ◽

10.1093/biolre/ioz111 ◽

2019 ◽

Vol 101 (3) ◽

pp. 567-578 ◽

Cited By ~ 1

Author(s):

Katsuhiko Hayashi

Keyword(s):

Germ Cell ◽

Cell Biology ◽

New Technology ◽

Rapid Development ◽

Cell Development ◽

Alternative Source ◽

Germ Cell Development ◽

In Vitro Reconstitution ◽

Key Issues

Abstract Germ cell development is a series of highly specialized processes through which diploid pluripotent cells differentiate into haploid gametes. The processes include biologically important events such as epigenetic reprogramming, sex determination, and meiosis. The mechanisms underlying these events are key issues in reproductive and developmental biology, yet they still remain elusive. As a tool to elucidate these mechanisms, in vitro gametogenesis, which reproduces germ cell development in culture, has long been sought for decades. Recently, methods of in vitro gametogenesis have undergone rapid development in association with stem cell biology, opening many possibilities in this field. This new technology is considered an alternative source of gametes for the reproduction of animals and perhaps humans. This review summarizes current advances and problems in in vitro gametogenesis.

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