Reprogramming of chimpanzee fibroblasts into a multipotent cancerous but not fully pluripotent state by transducing iPSC factors in 2i/LIF culture

Lin, Zachary Yu Ching; Nakai, Risako; Hirai, Hirohisa; Kozuka, Daiki; Katayama, Seiya; Nakamura, Shin; Okada, Sawako; Kitajima, Ryunosuke; Imai, Hiroo; Imamura, Makoto

doi:10.1016/j.diff.2020.01.002

Cited by 6 publications

(5 citation statements)

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“…Horse serum can increase the number of embryonic cells and promote the formation and hatching of blastocysts, which facilitates embryogenesis [54]. N2B27 medium can induce and sustain naïve pluripotency in mouse embryos [55].…”

Section: Discussionmentioning

confidence: 99%

A 3D"Sandwich" Co-culture System with hPPSCs and hUVECs Supports Mouse Embryo Development from E3.5 to E7.5 In Vitro

Teng

et al. 2023

Preprint

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Background A variety of methods for ex-utero culture systems have been explored, but there remain limitations in terms of in vitro culture platforms used prior to the implantation of mouse embryos, and the ability of mouse blastocysts to develop normally in vitro has not been established. Whether vascular niche support mouse embryo development from embryonic day (E) 3.5 to E7.5 in vitro is unknown.Methods We established a three-dimensional (3D) “sandwich” vascular-niche culture system with in vitro culture medium (IVCM) with human placenta perivascular stem cells (hPPSCs) and human umbilical vein endothelial cells (hUVECs), as supportive cells, which were seeded into the bottom layer of Matrigel to test mouse embryos from E3.5 to E7.5 in vitro. Mouse embryos from E3.5 to E7.5 development rates and greatest diameters at each stage were quantitatively determinated using statistics of SPSS software. Pluripotent markers and embryo transplantation in vivo were used to monitor mouse embryo quality and function.Results Embryos in the IVCM + Cells (hPPSCs + hUVECs) showed higher development rates and greatest diameters at each stage than those in the IVCM group. Embryos in the IVCM + Cells group cultured to E5.5 resembled natural egg cylinders in morphology and expressed specific embryonic cell markers, including Oct4 and Nanog, which were features similar to embryos developed in vivo. After transplantation, the embryos could be re-implanted in the internal uterus and continue to develop to a certain stage.Conclusions Therefore, the 3D in vitro culture system enabled the development of embryos from E3.5 to E7.5, and the vascularization microenvironment constructed by Matrigel, hPPSCs, and hUVECs significantly promoted the development of implanted embryos. This system allowed us to further study the physical and molecular mechanisms of embryo implantation in vitro.

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Section: Discussionmentioning

confidence: 99%

A 3D"Sandwich" Co-culture System with hPPSCs and hUVECs Supports Mouse Embryo Development from E3.5 to E7.5 In Vitro

Teng

et al. 2023

Preprint

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show abstract

“…Horse serum can increase the number of embryonic cells and promote the formation and hatching of blastocysts, which facilitates embryogenesis [ 56 ]. N2B27 medium can induce and sustain naïve pluripotency in mouse embryos [ 57 ].…”

Section: Discussionmentioning

confidence: 99%

A 3D “sandwich” co-culture system with vascular niche supports mouse embryo development from E3.5 to E7.5 in vitro

Xu,

Zhang,

et al. 2023

Stem Cell Res Ther

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Background Various methods for ex utero culture systems have been explored. However, limitations remain regarding the in vitro culture platforms used before implanting mouse embryos and the normal development of mouse blastocysts in vitro. Furthermore, vascular niche support during mouse embryo development from embryonic day (E) 3.5 to E7.5 is unknown in vitro. Methods This study established a three-dimensional (3D) “sandwich” vascular niche culture system with in vitro culture medium (IVCM) using human placenta perivascular stem cells (hPPSCs) and human umbilical vein endothelial cells (hUVECs) as supportive cells (which were seeded into the bottom layer of Matrigel) to test mouse embryos from E3.5 to E7.5 in vitro. The development rates and greatest diameters of mouse embryos from E3.5 to E7.5 were quantitatively determined using SPSS software statistics. Pluripotent markers and embryo transplantation were used to monitor mouse embryo quality and function in vivo. Results Embryos in the IVCM + Cells (hPPSCs + hUVECs) group showed higher development rates and greater diameters at each stage than those in the IVCM group. Embryos in the IVCM + Cells group cultured to E5.5 morphologically resembled natural egg cylinders and expressed specific embryonic cell markers, including Oct4 and Nanog. These features were similar to those of embryos developed in vivo. After transplantation, the embryos were re-implanted in the internal uterus and continued to develop to a particular stage. Conclusions The 3D in vitro culture system enabled embryo development from E3.5 to E7.5, and the vascularization microenvironment constructed by Matrigel, hPPSCs, and hUVECs significantly promoted the development of implanted embryos. This system allowed us to further study the physical and molecular mechanisms of embryo implantation in vitro.

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“…Beyond the pluripotent state, a growing number of studies has compared iPSC-differentiated cells from humans and other evolutionarily relevant species (Figure 2B), again considering both gene expression and regulation -although there are significantly fewer datasets available that speak to the latter. While the phenotypic difference between humans and other great apes that has drawn the most focus is the advanced cognitive skills that are unique to humans [113,114], which we discuss below, comparative studies have focused on an array of organs and cell types across all developmental tissue layers [101,109,112,[115][116][117][118]. Much of this work incorporates specific developmental questions or experimental manipulation that would be beyond the scope of non-iPSC models.…”

Section: Accepted Articlementioning

confidence: 99%

“…For instance, multiple groups have described interspecies differences in gene expression and regulation at the pluripotent stage that could have bearing on adult phenotypes [95,103,104,[109][110][111][112]. Marchetto et al [95] reported differences in activity in LINE-1 retrotransposons between four human iPSC lines and two chimpanzee and two bonobo lines.…”

Section: Interspecies Insights From Ipscderived Models For Human Evol...mentioning

confidence: 99%

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Harnessing pluripotent stem cells as models to decipher human evolution

Dannemann

Romero

2021

The FEBS Journal

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The study of human evolution, long constrained by a lack of experimental model systems, has been transformed by the emergence of the induced pluripotent stem cell (iPSC) field. iPSCs can be readily established from non-invasive tissues sources, both from humans and other primates; they can be maintained in the laboratory indefinitely and they can be differentiated into other tissue types. These qualities mean that iPSCs are rapidly becoming established as viable and powerful model systems with which it is possible to address questions in human evolution that were until now logistically and ethically intractable, especially in the quest to understand humans' place amongst the great apes, and the genetic basis of human uniqueness. In this review, we discuss the key lessons and takeaways of this nascent field; from the types of research iPSCs make possible to lingering challenges and likely future directions. We provide a comprehensive overview of how the seemingly unlikely combination of iPSCs and explicit evolutionary frameworks are transforming what is possible in our understanding of humanity's past and present.

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Reprogramming of chimpanzee fibroblasts into a multipotent cancerous but not fully pluripotent state by transducing iPSC factors in 2i/LIF culture

Cited by 6 publications

References 44 publications

A 3D"Sandwich" Co-culture System with hPPSCs and hUVECs Supports Mouse Embryo Development from E3.5 to E7.5 In Vitro

A 3D"Sandwich" Co-culture System with hPPSCs and hUVECs Supports Mouse Embryo Development from E3.5 to E7.5 In Vitro

A 3D “sandwich” co-culture system with vascular niche supports mouse embryo development from E3.5 to E7.5 in vitro

Harnessing pluripotent stem cells as models to decipher human evolution

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