scholarly journals Robust In Vitro Induction of Human Germ Cell Fate from Pluripotent Stem Cells

2015 ◽  
Vol 17 (2) ◽  
pp. 178-194 ◽  
Author(s):  
Kotaro Sasaki ◽  
Shihori Yokobayashi ◽  
Tomonori Nakamura ◽  
Ikuhiro Okamoto ◽  
Yukihiro Yabuta ◽  
...  
Keyword(s):  
Stem Cells ◽  
Germ Cell ◽  
Cell Fate ◽  
Pluripotent Stem Cells ◽  
In Vitro Induction

scholarly journals Simulating gastrulation development and germ cell fate in vitro using human and monkey pluripotent stem cells

2017 ◽  
Author(s):  
Toshihiro Kobayashi ◽  
Toshihiro Kobayashi ◽  
Ramiro Alberio ◽  
M Azim Surani
Keyword(s):  
Stem Cells ◽  
Germ Cell ◽  
Cell Fate ◽  
Pluripotent Stem Cells

scholarly journals Efficient generation of human primordial germ cell-like cells from pluripotent stem cells in a methylcellulose-based 3D system at large scale

PeerJ ◽  
2019 ◽  
Vol 6 ◽  
pp. e6143 ◽  
Author(s):  
Xiaoman Wang ◽  
Tingting Liao ◽  
Cong Wan ◽  
Xiaoyu Yang ◽  
Jiexiang Zhao ◽  
...  
Keyword(s):  
Gene Expression ◽  
Stem Cells ◽  
Germ Cell ◽  
Induced Pluripotent Stem Cells ◽  
Pluripotent Stem Cells ◽  
Large Scale ◽  
Induction System ◽  
Induced Pluripotent ◽  
In Vitro Induction

Background The mechanisms underlying human germ cell development and infertility remain largely unknown due to bioethical issues and the shortage of experimental materials. Therefore, an effective in vitro induction system of human primordial germ-like cells (hPGCLCs) from human pluripotent stem cells (hPSC) is in high demand. The current strategies used for the generation of hPGCLCs are not only costly but also difficult to perform at a large scale, thereby posing barriers to further research. In this study, we attempted to solve these problems by providing a new 3D culture system for hPGCLC differentiation. Methods The efficiency and relative yield of a methylcellulose (MC)-based 3D hPGCLC induction system were first compared with that of a conventional U96 system. Then, we examined the gene expression of germ cell marker genes and the key epigenetic modifications of the EpCAM-/INTEGRINα6-high cells from the 3D MC induction system and the U96 system via quantitative PCR and immunofluorescence. Finally, the reliability of the MC-based 3D hPGCLC induction system was evaluated via the generation of induced pluripotent stem cells (iPSCs) from the testicular cells of one patient with obstructive azoospermia (OA) and followed by the subsequent differentiation of iPSCs into the germ cell lineage. Results In the present study, we demonstrated that the 3D MC induction system combined with low-cell attachment plates facilitated the generation of hPGCLCs at a large scale. We found that the hPGCLCs generated via the MC system shared similar characteristics to that via the U96 system in terms of the gene expression profiles, germ cell-specific markers, epigenetic modification states and cellular states. In addition, hPGCLCs from iPSCs derived from one OA patient were generated with high efficiency via the present 3D MC induction system. Discussion The in vitro induction of hPGCLCs from human embryonic stem cells (hESCs)/human induced pluripotent stem cells (hiPSCs) has significant implications in exploring the underlying mechanisms of the origin and specification of hPGCs and the epigenetic programming of the human germ line as well as treating male infertility. Here, we developed a simple and efficient 3D induction system to generate hPGCLCs from hESCs/hiPSCs at a large scale, which facilitated the study of human germ cell development and stem cell-based reproductive medicine.

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

2019 ◽  
Vol 102 (3) ◽  
pp. 620-638 ◽  
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.

scholarly journals In Vitro Induction and In Vivo Engraftment of Lung Bud Tip Progenitor Cells Derived from Human Pluripotent Stem Cells

2018 ◽  
Vol 10 (1) ◽  
pp. 101-119 ◽  
Author(s):  
Alyssa J. Miller ◽  
David R. Hill ◽  
Melinda S. Nagy ◽  
Yoshiro Aoki ◽  
Briana R. Dye ◽  
...  
Keyword(s):  
Stem Cells ◽  
Progenitor Cells ◽  
Pluripotent Stem Cells ◽  
Human Pluripotent Stem Cells ◽  
In Vitro Induction

scholarly journals In vitro induction and in vivo engraftment of kidney organoids derived from human pluripotent stem cells

2020 ◽  
Vol 20 (2) ◽  
pp. 1307-1314
Author(s):  
Denglu Zhang ◽  
Xiaohang Du ◽  
Xufeng Zhang ◽  
Kailin Li ◽  
Feng Kong ◽  
...  
Keyword(s):  
Stem Cells ◽  
Pluripotent Stem Cells ◽  
Human Pluripotent Stem Cells ◽  
In Vitro Induction ◽  
Kidney Organoids

scholarly journals Cellular Programming and Reprogramming: Sculpting Cell Fate for the Production of Dopamine Neurons for Cell Therapy

2012 ◽  
Vol 2012 ◽  
pp. 1-17 ◽  
Author(s):  
Julio C. Aguila ◽  
Eva Hedlund ◽  
Rosario Sanchez-Pernaute
Keyword(s):  
Stem Cells ◽  
Cell Fate ◽  
Pluripotent Stem Cells ◽  
Dorsal Striatum ◽  
Dopamine Neurons ◽  
Cell Fates ◽  
Intrinsic Factors ◽  
Neuronal Identity ◽  
The Right

Pluripotent stem cells are regarded as a promising cell source to obtain human dopamine neurons in sufficient amounts and purity for cell replacement therapy. Importantly, the success of clinical applications depends on our ability to steer pluripotent stem cells towards the right neuronal identity. In Parkinson disease, the loss of dopamine neurons is more pronounced in the ventrolateral population that projects to the sensorimotor striatum. Because synapses are highly specific, only neurons with this precise identity will contribute, upon transplantation, to the synaptic reconstruction of the dorsal striatum. Thus, understanding the developmental cell program of the mesostriatal dopamine neurons is critical for the identification of the extrinsic signals and cell-intrinsic factors that instruct and, ultimately, determine cell identity. Here, we review how extrinsic signals and transcription factors act together during development to shape midbrain cell fates. Further, we discuss how these same factors can be appliedin vitroto induce, select, and reprogram cells to the mesostriatal dopamine fate.

scholarly journals Micropattern differentiation of mouse pluripotent stem cells recapitulates embryo regionalized cell fate patterning

eLife ◽  
2018 ◽  
Vol 7 ◽  
Author(s):  
Sophie M Morgani ◽  
Jakob J Metzger ◽  
Jennifer Nichols ◽  
Eric D Siggia ◽  
Anna-Katerina Hadjantonakis
Keyword(s):  
Stem Cells ◽  
Cell Fate ◽  
Pluripotent Stem Cells ◽  
Epithelial To Mesenchymal Transition ◽  
Cell Types ◽  
Mesenchymal Transition ◽  
Fate Specification ◽  
Epiblast Stem Cells

During gastrulation epiblast cells exit pluripotency as they specify and spatially arrange the three germ layers of the embryo. Similarly, human pluripotent stem cells (PSCs) undergo spatially organized fate specification on micropatterned surfaces. Since in vivo validation is not possible for the human, we developed a mouse PSC micropattern system and, with direct comparisons to mouse embryos, reveal the robust specification of distinct regional identities. BMP, WNT, ACTIVIN and FGF directed mouse epiblast-like cells to undergo an epithelial-to-mesenchymal transition and radially pattern posterior mesoderm fates. Conversely, WNT, ACTIVIN and FGF patterned anterior identities, including definitive endoderm. By contrast, epiblast stem cells, a developmentally advanced state, only specified anterior identities, but without patterning. The mouse micropattern system offers a robust scalable method to generate regionalized cell types present in vivo, resolve how signals promote distinct identities and generate patterns, and compare mechanisms operating in vivo and in vitro and across species.

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