Improving Cell Survival in Injected Embryos Allows Primed Pluripotent Stem Cells to Generate Chimeric Cynomolgus Monkeys

Kang, Yu; Ai, Zongyong; Duan, Kui; Si, Chenyang; Wang, Yong; Zheng, Yun; He, Jingjing; Yin, Yu; Zhao, Shumei; Niu, Ben; Zhu, Xiaoqing; Liu, Li; Xiang, Lifeng; Zhang, Linming; Niu, Yuyu; Ji, Weizhi; Li, Tianqing

doi:10.1016/j.celrep.2018.11.001

Cited by 22 publications

(17 citation statements)

References 52 publications

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“…Although it is ambiguous whether the STB-like syncytia are derived through the progenitor state corresponding to embryonic CTBs, these results indicate that human pESCs possess the competence to give rise to not only STBs in primitive syncytium of the implantation embryo but also an in vitro counterpart of villous CTBs in the firsttrimester placenta. In a previous report, monkey pESCs were shown to contribute to the placental tissue when injected into the morula stage of the monkey embryo, supporting our results (Kang et al, 2018).…”

Section: Discussionsupporting

confidence: 92%

Cell-autonomous differentiation of human primed embryonic stem cells into trophoblastic syncytia through the nascent amnion-like cell state

Ohgushi

Eiraku

2021

Preprint

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Human primed embryonic stem cells (pESCs) are known to be converted to cells with several trophoblast properties, but it has remained controversial whether this phenomenon represents the inherent differentiation competence of human pESCs to trophoblast lineages. In this study, we report that chemical blockage of ACTIVIN/NODAL and FGF signals is sufficient to steer human pESCs into GATA3-expressing cells that give rise to hormone-producing syncytia analogous to syncytiotrophoblasts of the post-implantation stage of the human embryo. Taking advantage of this system, we identified two distinct modes of cell-autonomous genetic programs and their coordinated actions to initiate the differentiation. We also found a transient population reminiscent of nascent amnion and then a spontaneous branch of differentiation trajectory leading to syncytiotrophoblast-like syncytial cells. These results provide insights into the possible extraembryonic differentiation pathway that is unique in primate embryogenesis and is relevant to the trophoblast competence of human primed pluripotent stem cells.

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Section: Discussionsupporting

confidence: 92%

Cell-autonomous differentiation of human primed embryonic stem cells into trophoblastic syncytia through the nascent amnion-like cell state

Ohgushi

Eiraku

2021

Preprint

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“…Primate ESCs exist in a different phase of pluripotency than rodent ESCs, as they are primed rather than naïve (Brons et al, 2007;Nichols and Smith, 2009; Boroviak et al, 2014). The ratio of cmESCs in allogenic chimeras is only 0.1%-4.5% (Chen et al, 2015;Kang et al, 2018), which is significantly lower than with rodents. However, in previous reports, naïve hPSCs have lacked the competency to integrate into the mouse embryo (Gafni et al, 2013;Theunissen et al, 2014).…”

Section: Discussionmentioning

confidence: 97%

“…In our domestication system, the cell and embryo culture media were mixed in an optimal ratio, increasing the anti-apoptotic ability of cmESCs and improving the development of pig embryos, and allowing the successful generation of chimeric animals. Improvement of the culture conditions also played an important role in a previous homozygous chimera study (Kang et al, 2018).…”

Section: Discussionmentioning

confidence: 99%

“…Although primed hPSCs could not participate in normal mouse development when transplanted into preimplantation embryos as naïve PSCs (Mascetti and Pedersen, 2016), inhibiting apoptosis was shown to enhance the chimeric ability of primed hPSCs in mice (Huang et al, 2018;Wang et al, 2018b). In addition, Kang et al showed that primed PSCs from cynomolgus monkeys (Macaca fascicularis) could generate chimeras using optimal culture conditions (Kang et al, 2018). Several recent studies have also reported the generation of naive PSCs, which showed higher efficiency in single-cell cloning and genome editing, and could be incorporated in the host when transplanted into preimplantation embryos (Gafni et al, 2013;Chen et al, 2015).…”

Section: Introductionmentioning

confidence: 99%

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Domesticated cynomolgus monkey embryonic stem cells allow the generation of neonatal interspecies chimeric pigs

Ren

et al. 2019

Protein Cell

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Blastocyst complementation by pluripotent stem cell (PSC) injection is believed to be the most promising method to generate xenogeneic organs. However, ethical issues prevent the study of human chimeras in the late embryonic stage of development. Primate embryonic stem cells (ESCs), which have similar pluripotency to human ESCs, are a good model for studying interspecies chimerism and organ generation. However, whether primate ESCs can be used in xenogenous grafts remains unclear. In this study, we evaluated the chimeric ability of cynomolgus monkey (Macaca fascicularis) ESCs (cmESCs) in pigs, which are excellent hosts because of their many similarities to humans. We report an optimized culture medium that enhanced the anti-apoptotic ability of cmESCs and improved the development of chimeric embryos, in which domesticated cmESCs (D-ESCs) injected into pig blastocysts differentiated into cells of all three germ layers. In addition, we obtained two neonatal interspecies chimeras, in which we observed tissue-specific D-ESC differentiation. Taken together, the results demonstrate the capability of D-ESCs to integrate and differentiate into functional cells in a porcine model, with a chimeric ratio of 0.001–0.0001 in different neonate tissues. We believe this work will facilitate future developments in xenogeneic organogenesis, bringing us one step closer to producing tissue-specific functional cells and organs in a large animal model through interspecies blastocyst complementation.

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“…Previous studies have shown that inhibiting apoptosis either by the addition of the Rock inhibitor Y27632 (Kang et al, 2018), or by overexpression of the anti-apoptotic genes BCL2 (Masaki et al, 2016) or BMI1 (Huang et al, 2018) enables primed PSCs to colonize host embryos. Therefore, in a final series of microinjections, we asked whether inhibition of apoptosis could improve the engraftment of naïve PSCs into the host embryo.…”

Section: Premature Differentiation Of Rhesus Naïve Pscs After Injectimentioning

confidence: 99%

Apoptosis, G1 phase stall and premature differentiation account for low chimeric competence of Human and rhesus monkey naïve pluripotent stem cells

Aksoy

Rognard

Moulin

et al. 2020

Preprint

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After reprogramming to naïve pluripotency, human pluripotent stem cells (PSCs) still exhibit very low ability to make interspecies chimeras in mice and pigs. Whether this is because they are inherently devoid of the attributes of chimeric competency or because naïve PSCs in general cannot colonize embryos from distant species remains to be elucidated. Here, we have used different types of mouse, human and rhesus monkey naïve PSCs and we have analyzed their ability to colonize both distant (rabbit) and close (cynomolgus monkey) embryos. Mouse embryonic stem cells (ESCs) remained mitotically active after injection into rabbit and cynomolgus embryos and contributed to the formation of the neural tube in postimplantation rabbit embryos. In contrast, primate naive PSCs colonized rabbit and cynomolgus embryos with much lower efficiency, regardless of the reprogramming protocols. Unlike mouse ESCs, they slowed DNA replication following colony dissociation and, after injection into host embryos, they stalled in the G1 phase of the cell cycle and differentiated prematurely, regardless of host species, distant or close. These results show that, unlike mouse ESCs, primate naive PSCs do not make chimeras because they are inherently unfit to stay mitotically active during embryo colonization.

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Improving Cell Survival in Injected Embryos Allows Primed Pluripotent Stem Cells to Generate Chimeric Cynomolgus Monkeys

Cited by 22 publications

References 52 publications

Cell-autonomous differentiation of human primed embryonic stem cells into trophoblastic syncytia through the nascent amnion-like cell state

Cell-autonomous differentiation of human primed embryonic stem cells into trophoblastic syncytia through the nascent amnion-like cell state

Domesticated cynomolgus monkey embryonic stem cells allow the generation of neonatal interspecies chimeric pigs

Apoptosis, G1 phase stall and premature differentiation account for low chimeric competence of Human and rhesus monkey naïve pluripotent stem cells

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