Suppression of ES cell differentiation by retinol (vitamin A) via the overexpression of Nanog

Chen, Liguo; Yang, Mengjin; Dawes, Joyce; Khillan, Jaspal S.

doi:10.1111/j.1432-0436.2007.00169.x

Cited by 35 publications

(43 citation statements)

References 44 publications

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“…Retinoic acid, one of the retinoids, plays an essential role in early development and maintenance of specific tissue types. Khillan and colleagues demonstrated that retinol can be used to maintain mESCs in a pluripotent state in feeder-free cultures (Chen et al., 2007). Retinol increases the expression of NANOG and promotes self-renewal by activating PI3K/AKT and the IGF-1/IRS-1 pathway (Chen and Khillan, 2010; Chen et al., 2007).…”

Section: Discussionmentioning

confidence: 99%

A Method to Identify and Isolate Pluripotent Human Stem Cells and Mouse Epiblast Stem Cells Using Lipid Body-Associated Retinyl Ester Fluorescence

et al. 2014

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SummaryWe describe the use of a characteristic blue fluorescence to identify and isolate pluripotent human embryonic stem cells and human-induced pluripotent stem cells. The blue fluorescence emission (450–500 nm) is readily observed by fluorescence microscopy and correlates with the expression of pluripotency markers (OCT4, SOX2, and NANOG). It allows easy identification and isolation of undifferentiated human pluripotent stem cells, high-throughput fluorescence sorting and subsequent propagation. The fluorescence appears early during somatic reprogramming. We show that the blue fluorescence arises from the sequestration of retinyl esters in cytoplasmic lipid bodies. The retinoid-sequestering lipid bodies are specific to human and mouse pluripotent stem cells of the primed or epiblast-like state and absent in naive mouse embryonic stem cells. Retinol, present in widely used stem cell culture media, is sequestered as retinyl ester specifically by primed pluripotent cells and also can induce the formation of these lipid bodies.

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Section: Discussionmentioning

confidence: 99%

A Method to Identify and Isolate Pluripotent Human Stem Cells and Mouse Epiblast Stem Cells Using Lipid Body-Associated Retinyl Ester Fluorescence

et al. 2014

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“…Also, work by others suggests vitamin A is a valuable component to the sternness and growth of mouse ESCs. RA-independent and possibly RA-dependent roles for vitamin A are important for maintaining rates of mouse ESC proliferation and pluripotency (Chen et al, 2007; Chen and Khillan, 2010; Chen and Khillan, 2008; Khillan, 2014; Tagliaferri et al, 2016; Yang et al, 2015). Therefore, maintaining mouse ESCs in the absence of vitamin A is not likely a viable option, but could interfere with efficiency of subsequent differentiation steps as our studies possibly suggest.…”

Section: Discussionmentioning

confidence: 99%

Inverse agonism of retinoic acid receptors directs epiblast cells into the paraxial mesoderm lineage

Russell

Liu

et al. 2018

Stem Cell Research

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We have investigated the differentiation of paraxial mesoderm from mouse embryonic stem cells utilizing a Tbx6-EYFP/Brachyury (T)-Ckerry dual reporter system. Differentiation from the mouse ESC state directly into mesoderm via Wnt pathway activation was low, but augmented by treatment with AGN193109, a pan-retinoic acid receptor inverse agonist. After five days of differentiation, T+ cells increased from 12.2% to 18.8%, Tbx6+ cells increased from 5.8% to 12.7%, and T+/Tbx6+ cells increased from 2.4% to 14.1%. The synergism of AGN193109 with Wnt3a/CHIR99021 was further substantiated by the increased expression of paraxial mesoderm gene markers Tbx6, Msgnl, Meoxl, and Hoxbl. Separate to inverse agonist treatment, when mouse ESCs were indirectly differentiated into mesoderm via a transient epiblast step the efficiency of paraxial mesoderm formation markedly increased. Tbx6+ cells represented 65–75% of the total cell population after just 3 days of differentiation and the expression of paraxial mesoderm marker genes Tbx6 and Msgn increased over 100-fold and 300-fold, respectively. Further evaluation of AGN193109 treatment on the indirect differentiation protocol suggested that RARs have two distinct roles. First, AGN193109 treatment at the epiblast step and mesoderm step promoted paraxial mesoderm formation over other mesoderm and endoderm lineage types. Second, continued treatment during mesoderm formation revealed its ability to repress the maturation of presomitic mesoderm into somitic paraxial mesoderm. Thus, the continuous treatment of AGN193109 during epiblast and mesoderm differentiation steps yielded a culture where —90% of the cells were Tbx6+. The surprisingly early effect of inverse agonist treatment at the epiblast step of differentiation led us to further examine the effect of AGN193109 treatment during an extended epiblast differentiation protocol. Interestingly, while inverse agonist treatment had no impact on the conversion of ESCs into epiblast cells based on the expression of Rexl, Fgf5, and pluripotency marker genes Oct4, Nanog, and Sox2, after three days of differentiation in the presence of AGN193109 caudal epiblast and early paraxial mesoderm marker genes, T, Cyp26al, Fgf8, Tbx6 and Msgn were all highly up-regulated. Collectively, our studies reveal an earlier than appreciated role for RARs in epiblast cells and the modulation of their function via inverse agonist treatment can promote their differentiation into the paraxial mesoderm lineage.

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“…Several other transcription factors, including Sall4, FoxD3, p53, and Tcf3, have been implicated in regulating Nanog gene expression (Cole et al, 2008;Hanna et al, 2002;Lim et al, 2008;Liu and Labosky, 2008;Pan and Thomson, 2007;Pereira et al, 2006;Wu et al, 2006). Signal-wise, Nanog expression levels have been reported to be modulated in various systems by LIF, Wnt, BMP, FGF, retinol and Activin/Nodal TGF-beta pathways (Boiani and Scholer, 2005;Chen et al, 2007Chen et al, , 2008Kunath et al, 2007;Okita and Yamanaka, 2006;Sato et al, 2004;Silva et al, 2008;Suzuki et al, 2006;Takao et al, 2007;Vallier et al, 2009;Xu et al, 2008;Yi et al, 2008;Ying et al, 2008). Despite these in vitro studies, the regulation of Nanog expression in vivo is not well understood.…”

Section: Introductionmentioning

confidence: 99%

Activin/TGF-beta signaling regulates Nanog expression in the epiblast during gastrulation

Shin¹,

Alev²,

Wu³

et al. 2011

Mechanisms of Development

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Nanog is required for the maintenance of cellular pluripotency during normal development and in cultured embryonic stem cells. A number of signaling pathways have been implicated in regulating Nanog gene expression in vitro. Using the chick model, we provide in vivo evidence for the involvement of the Activin/TGF-beta signaling pathway in regulating Nanog expression in epiblast cells during gastrulation. Nanog expression in primordial germ cells is not regulated by this pathway, indicating that these two cell types employ different mechanisms for maintaining pluripotency in early development. Furthermore, our data suggest that the bHLH factor E2A plays a role in negatively regulating Nanog expression in vivo. Overall, our data support a direct and positive role of the Smad2/3 mediated TGF-beta signaling pathway in inducing/maintaining Nanog expression.

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Suppression of ES cell differentiation by retinol (vitamin A) via the overexpression of Nanog

Cited by 35 publications

References 44 publications

A Method to Identify and Isolate Pluripotent Human Stem Cells and Mouse Epiblast Stem Cells Using Lipid Body-Associated Retinyl Ester Fluorescence

A Method to Identify and Isolate Pluripotent Human Stem Cells and Mouse Epiblast Stem Cells Using Lipid Body-Associated Retinyl Ester Fluorescence

Inverse agonism of retinoic acid receptors directs epiblast cells into the paraxial mesoderm lineage

Activin/TGF-beta signaling regulates Nanog expression in the epiblast during gastrulation

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