Insulin fine-tunes self-renewal pathways governing naive pluripotency and extra-embryonic endoderm

Anderson, Kathryn G.; Hamilton, William B.; Roske, Fabian V; Azad, Ajuna; Knudsen, Teresa E.; Canham, Maurice A.; Forrester, Lesley M.; Brickman, Joshua M.

doi:10.1038/ncb3617

Cited by 75 publications

(125 citation statements)

References 64 publications

Supporting

Mentioning

111

Contrasting

Order By: Relevance

“…EBs that formed from pXENCs exhibited a more irregular morphology than those derived from piPSCs ( Figure 3A RA induces differentiation of XEN cells. 23 In culture, pXENCs exhibited dispersed morphology ( Figure 3D) but became highly aggregated after the addition of RA ( Figure 3E). After RA addition, the levels of XEN markers, such as Gata4, Gata6, Sox17 and Pdgfra, remained unchanged compared with those in the control group that without RA addition.…”

Section: Differentiation Potential Of Pxencsmentioning

confidence: 98%

Characterization of porcine extraembryonic endoderm cells

Shen

Zhang

et al. 2019

Cell Proliferation

View full text Add to dashboard Cite

Objectives To date, many efforts have been made to establish porcine embryonic stem (pES) cells without success. Extraembryonic endoderm (XEN) cells can self‐renew and differentiate into the visceral endoderm and parietal endoderm. XEN cells are derived from the primitive endoderm of the inner cell mass of blastocysts and may be an intermediate state in cell reprogramming. Materials and methods Porcine XEN cells (pXENCs) were generated from porcine pluripotent stem cells (pPSCs) and were characterized by RNA sequencing and immunofluorescence analyses. The developmental potential of pXENCs was investigated in chimeric mouse embryos. Results Porcine XEN cells derived from porcine pPSCs were successfully expanded in N2B27 medium supplemented with bFGF for least 30 passages. RNA sequencing and immunofluorescence analyses showed that pXENCs expressed the murine and canine XEN markers Gata6 , Gata4 , Sox17 and Pdgfra but not the pluripotent markers Oct4 , Sox2 and TE marker Cdx2 . Moreover, these cells contributed to the XEN when injected into four‐cell stage mouse embryos. Supplementation with Chir99021 and SB431542 promoted the pluripotency of the pXENCs. Conclusions We successfully derived pXENCs and showed that supplementation with Chir99021 and SB431542 confer them with pluripotency. Our results provide a new resource for investigating the reprogramming mechanism of porcine‐induced pluripotent stem cells.

show abstract

Section: Differentiation Potential Of Pxencsmentioning

confidence: 98%

Characterization of porcine extraembryonic endoderm cells

Shen

Zhang

et al. 2019

Cell Proliferation

View full text Add to dashboard Cite

show abstract

“…This is not unexpected. Wnt and (Activin-like) Nodal epithelialize XEN and pXEN cells (Bangs, Schrode, Hadjantonakis, & Anderson, 2015;Chuykin et al, 2013;Kruithof-de Julio et al, 2011), and nEnd cells are epithelial and cultured with Activin and Wnt pathway activation (Anderson et al, 2017). However, Tgf-β had not been invoked in the PE phenotype, although it is a key mesenchyme regulator (Moustakas & Heldin, 2016) and present in serum (Danielpour et al, 1989), which is required for XEN and pXEN cells (Supporting Information Table S1).…”

Section: Discussionmentioning

confidence: 99%

“…The first group includes mouse blastocyst‐derived ExEn stem cell lines named XEN cells (Kunath et al, ) and many similar lines produced from ES cells, yolk sacs, or (by reprogramming) fibroblasts (e.g., Cho et al, ; Lin, Khan, Zapiec, & Mombaerts, ; Parenti, Halbisen, Wang, Latham, & Ralston, ; Shimosato, Shiki, & Niwa, ). The second group includes rat and mouse blastocyst‐derived “primitive XEN (pXEN)” (Debeb et al, ; Zhong et al, ) and mouse ES cell‐derived “naïve extraembryonic endoderm progenitor (nEnd)” (Anderson et al, ) cell lines. In addition, numerous PrE‐related cells were isolated from ES cell cultures but not grown as lines (e.g., Canham, Sharov, Ko, & Brickman, ; Lo Nigro et al, ); from this group, we selected the “PrE‐primed ES cells” (Lo Nigro et al, ).…”

Section: Introductionmentioning

confidence: 99%

“…In addition, numerous PrE‐related cells were isolated from ES cell cultures but not grown as lines (e.g., Canham, Sharov, Ko, & Brickman, ; Lo Nigro et al, ); from this group, we selected the “PrE‐primed ES cells” (Lo Nigro et al, ). The second group and PrE‐like ES cells are distinct from the “XEN group” as follows: They require the cytokine LIF (see also Supporting Information Table S1), express Pou5f1, can directly contribute to the VE and appeared closer to the blastocyst in transcriptome comparisons (Anderson et al, ; Debeb et al, ; Lo Nigro et al, ; Zhong et al, ). nEnd and pXEN cells were easily converted into XEN‐like cells, but not vice versa (Anderson et al, ; Zhong et al, ), again arguing that XEN cells are more differentiated.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Transcriptome analysis shows ambiguous phenotypes of murine primitive endoderm‐related stem cell lines

Zhong

Binas

2019

Genes to Cells

View full text Add to dashboard Cite

Primitive endoderm (PrE)‐related cell lines (XEN, pXEN and nEnd cells) show key features of the PrE. By transcriptome analysis, we show: (a) Compared to embryonic stem cells, PrE‐related cell lines are less in vivo like, although early nEnd cells are most similar to the PrE. (b) These cell lines show post‐PrE features of parietal (XEN and pXEN cells) or visceral (nEnd cells) endoderm, likely driven by Tgf‐β and Wnt/Activin signaling, respectively. (c) pXEN and nEnd cell lines additionally show pre‐PrE features. Hence, neither pXEN nor nEnd cell cultures represent a distinct in vivo entity. Rather, their properties are compatible with mixed and hybrid phenotypes. Our findings indicate that pre‐PrE, PrE and early post‐PrE phenotypes result from different niches, which need to be better understood to derive cell lines that distinctly represent the early stages of the extraembryonic endoderm.

show abstract

“…The XEN cells can also be induced from ESC by overexpression of PrEspecific genes, Gata-4, 6(6, 7), or Sox17 (8), or by treatment with growth factors (9). More recently, naïve extraembryonic endodermal cells (nEnd) resembling the blastocyst-stage PrE-precursors have been developed from the authentic mouse ESC (10). In rat, XEN cells established from blastocysts have different culture requirements and gene expression profiles compared to mouse XEN cells (11,12).…”

Section: Introductionmentioning

confidence: 99%

Chimera-competent eXtra-Embryonic eNdoderm (XEN) cells established from pig embryos

Park

Jeoung

et al. 2020

Preprint

View full text Add to dashboard Cite

Classification Biological Sciences/ Agricultural Sciences Keywords primitive endoderm, stem cell, chimera, XEN, SCNT Author Contributions CP, JCB and BT conceived the project and designed the experiments; CP, YJ, YU and JB established and performed characterization of embryonic outgrowths and XEN cells; KP and AP performed the knockin and generated the GFP reporter lines used for generating GFP:XEN cells used in chiera experiments; CP, JL, LP, TL, JS, YG, YS, JW 2 generated the libraries and transcriptomic analysis; CP and BT wrote the initial draft, CP, JW, JCB and BT revised the manuscript based on the input from all authors. All authors approved final draft for submission. Abstract In this article, we report for the first time the derivation and characterization of extraembryonic endoderm (XEN) cells from primitive endoderm (PrE) of porcine (p) embryos. The pXEN cells can be reliably and reproducibly generated from parthenote, in vitro and in vivo derived embryos. The pXEN cells retained all the hallmarks of PrE including expression of canonical PrE and XEN cell markers (GATA4, GATA6, SOX17, SALL4, FOXA2, and HNF4A). Transcriptome analysis further confirmed their XEN cell origin. The pXEN cells when introduced into blastocyst stage embryo contributed to widespread chimerism including visceral yolk sac, chorion, as well as embryonic gut and liver primordium in the fetus. The pXEN cells were shown to be an efficient nuclear donor for generating cloned offspring. Taken together, pXEN cells fulfil a longstanding need for a stable, chimera-competent, and nuclear transfer-compatible porcine embryonic cells with applications for agriculture and medicine. Significance StatementWe report for the first time, the derivation and characterization of extraembryonic endoderm (XEN) stem cells from porcine (p) embryos. The pXEN cells can be reliably and reproducibly derived from primitive endoderm precursors. When injected into blastocyst-stage embryos, the pXEN cells have contributed to wide-spread chimerism including visceral yolk sac, chorion of the extraembryonic membranes, as well as definitive endoderm of the fetus, primarily the embryonic gut and liver primordium.Additionally, these XEN cells have proven to be an efficient nuclear donor for generating cloned offspring. These newly discovered stem cells provide a novel model for studying 4 lineage segregation, as well as a source for interspecies chimeras for generating endodermal organs, and for genome editing in livestock. 5

show abstract

Insulin fine-tunes self-renewal pathways governing naive pluripotency and extra-embryonic endoderm

Cited by 75 publications

References 64 publications

Characterization of porcine extraembryonic endoderm cells

Characterization of porcine extraembryonic endoderm cells

Transcriptome analysis shows ambiguous phenotypes of murine primitive endoderm‐related stem cell lines

Chimera-competent eXtra-Embryonic eNdoderm (XEN) cells established from pig embryos

Contact Info

Product

Resources

About