Tuning of β-catenin activity is required to stabilize self-renewal of rat embryonic stem cells

Meek, Stephen; Wei, Jun; Sutherland, Linda; Nilges, Benedikt S.; Buehr, Mia; Tomlinson, Simon R.; Thomson, Axel A.; Burdon, Tom

doi:10.1002/stem.1466

Cited by 40 publications

(37 citation statements)

References 43 publications

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“…We note that the amount of GSK3i used influenced culture stability such that cells selfrenewed in 0-2 μM GSK3i, whereas 5 μM drove differentiation. This varies from mESCs, where 3 μM GSK3i is optimal (3), but is similar to observations made with naïve rat ESCs, for which 1 μM GSK3i is optimal (22,23). Excess GSK3i may enable Wnt-induced nuclear events that are counter to the naïve state.…”

Section: Discussionsupporting

confidence: 65%

Derivation of naïve human embryonic stem cells

Ware¹,

Nelson

Mecham

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

417

447

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The naïve pluripotent state has been shown in mice to lead to broad and more robust developmental potential relative to primed mouse epiblast cells. The human naïve ES cell state has eluded derivation without the use of transgenes, and forced expression of OCT4, KLF4, and KLF2 allows maintenance of human cells in a naïve state [Hanna J, et al. (2010) Proc Natl Acad Sci USA 107 (20):9222-9227]. We describe two routes to generate nontransgenic naïve human ES cells (hESCs). The first is by reverse toggling of preexisting primed hESC lines by preculture in the histone deacetylase inhibitors butyrate and suberoylanilide hydroxamic acid, followed by culture in MEK/ERK and GSK3 inhibitors (2i) with FGF2. The second route is by direct derivation from a human embryo in 2i with FGF2. We show that human naïve cells meet mouse criteria for the naïve state by growth characteristics, antibody labeling profile, gene expression, X-inactivation profile, mitochondrial morphology, microRNA profile and development in the context of teratomas. hESCs can exist in a naïve state without the need for transgenes. Direct derivation is an elusive, but attainable, process, leading to cells at the earliest stage of in vitro pluripotency described for humans. Reverse toggling of primed cells to naïve is efficient and reproducible.

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Section: Discussionsupporting

confidence: 65%

Derivation of naïve human embryonic stem cells

Ware¹,

Nelson

Mecham

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

417

447

View full text Add to dashboard Cite

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“…We supposed that this phenomenon was due to use of the GSK3 inhibitor, CHIR99021, during rPSC cultivation. This is in agreement with previous studies [33][34][35] that demonstrated that GSK3 inhibition led to activation of some differentiationassociated genes, including several trophoblast-specific genes. Similar to rPSCs, mouse ESCs and iPSCs were recently discovered to give rise to trophoblast cells due to Cdx2 overexpression [43], indicating the existence of common transcriptional mechanisms that regulate lineagespecific differentiation in rats and mice.…”

Section: Discussionsupporting

confidence: 93%

“…It was described that use of a GSK3 inhibitor, CHIR99021, in cultivation of rat and mouse ESCs caused ectopic expression of Cdx2 [33][34][35] encoding a wellknown trophoectodermal transcription factor [36]. According to RT-PCR and transcriptome analysis, Cdx2 was not expressed in the rEF lines and was highly expressed (on the average, 1,192 normalized read counts) in the rat pluripotent cells (Fig.…”

Section: Transcriptome Analysismentioning

confidence: 98%

Transcriptome Characteristics and X-Chromosome Inactivation Status in Cultured Rat Pluripotent Stem Cells

Vaskova

Medvedev

Sorokina

et al. 2015

Stem Cells and Development

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Rat pluripotent stem cells, embryonic stem cells (ESCs), and induced pluripotent stem cells (iPSCs) as mouse and human ones have a great potential for studying mammalian early development, disease modeling, and evaluation of regenerative medicine approaches. However, data on pluripotency realization and self-renewal maintenance in rat cells are still very limited, and differentiation protocols of rat ESCs (rESCs) and iPSCs to study development and obtain specific cell types for biomedical applications are poorly developed. In this study, the RNA-Seq technique was first used for detailed transcriptome characterization in rat pluripotent cells. The rESC and iPSC transcriptomes demonstrated a high similarity and were significantly different from those in differentiated cells. Additionally, we have shown that reprogramming of rat somatic cells to a pluripotent state was accompanied by X-chromosome reactivation. There were two active X chromosomes in XX rESCs and iPSCs, which is one of the key attributes of the pluripotent state. Differentiation of both rESCs and iPSCs led to X-chromosome inactivation (XCI). The dynamics of XCI in differentiating rat cells was very similar to that in mice. Two types of facultative heterochromatin described in various mammalian species were revealed on the rat inactive X chromosome. To explore XCI dynamics, we established a new monolayer differentiation protocol for rESCs and iPSCs that may be applied to study different biological processes and optimized for directed derivation of specific cell types.

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“…Importantly, we demonstrated that GRP78 downregulation was accompanied with the decreased expression of c-Myc and β-catenin (both are well characterized oncogene and stemness markers). Importantly, c-Myc has been characterized as one of the key factors for the maintenance of stem cell pluripotency and generation of cancer stem cells [31]; β-catenin, in addition, also has been indicated in different cancer types as a powerful oncogene and factor for stabilizing self-renewal ability [32,33]. The concomitant decreased expression of both c-Myc and β-catenin as the result of GRP78 down-regulation in GBM cells suggests a novel interaction between these molecules.…”

Section: Discussionmentioning

confidence: 99%