Derivation of Pre-X Inactivation Human Embryonic Stem Cells under Physiological Oxygen Concentrations

Lengner, Christopher J.; Gimelbrant, Alexander A.; Erwin, Jennifer A.; Cheng, Albert W.; Guenther, Matthew G.; Welstead, G. Grant; Alagappan, Raaji; Frampton, Garrett M.; Xu, Ping; Muffat, Julien; Santagata, Sandro; Powers, Doug; Barrett, C.B.; Young, Richard A.; Lee, Jeannie T.; Jaenisch, Rudolf; Mitalipova, Maisam

doi:10.1016/j.cell.2010.04.010

Cited by 368 publications

(366 citation statements)

References 74 publications

(73 reference statements)

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“…Identification of both Xs via XIST-FISH before X inactivation is consistent with a previous observation on human embryos (12). Thus, we looked at XIST promoter methylation as an indicator of an inactive X, using four primer sets (13). Sets 1L-1R, 9F-9R, and 11F-11R indicated similar extensive promoter methylation in the naïve, primed, and differentiated states.…”

Section: Elf1 Cellssupporting

confidence: 55%

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: Elf1 Cellssupporting

confidence: 55%

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

show abstract

“…Female hESCs derived from the www.cell-research.com | Cell Research Jia-Chi Yeo and Huck-Hui Ng 27 npg culture of human blastocysts in hypoxic (5% O 2 ) conditions were found to be in an XaXa status, a characteristic of mouse naive pluripotency [132]. However, these cells were maintained in conventional FGF-containing media and were not tested further for features of naive pluripotency.…”

Section: Mesc-like Hescsmentioning

confidence: 99%

The transcriptional regulation of pluripotency

Yeo

2012

Cell Res

114

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Abbreviations: 2C (2-cell); 2i (two inhibitor); BMP4 (bone morphogenic protein 4); ChIP (chromatin immunoprecipitation); ChIP-seq (chromatin immunoprecipitation and sequencing); ESCs (embryonic stem cells); EpiSCs (Epiblast-derived stem cells); Fgf4 (fibroblast growth factor 4); hESCs (human embryonic stem cells); ICM (inner cell mass); iPSCs (induced pluripotent stem cells); LIF (leukemia inhibitory factor); lincRNA (large intergenic non-coding RNA); mESCs (mouse embryonic stem cells); miRNA (micro RNA); ncRNA (non-coding RNA); PcG (Polycomb group proteins); PGCs (primodial germ cells); POU (Pit-Oct-Unc); RNAi (RNA interference); RNA-seq (RNA sequencing); SCF (stem cell factor); siRNA (short interfering RNA); XaXa (double X-chromosome active); XaXi (single X-chromosome inactivated)The defining features of embryonic stem cells (ESCs) are their self-renewing and pluripotent capacities. Indeed, the ability to give rise into all cell types within the organism not only allows ESCs to function as an ideal in vitro tool to study embryonic development, but also offers great therapeutic potential within the field of regenerative medicine. However, it is also this same remarkable developmental plasticity that makes the efficient control of ESC differentiation into the desired cell type very difficult. Therefore, in order to harness ESCs for clinical applications, a detailed understanding of the molecular and cellular mechanisms controlling ESC pluripotency and lineage commitment is necessary. In this respect, through a variety of transcriptomic approaches, ESC pluripotency has been found to be regulated by a system of ESC-associated transcription factors; and the external signalling environment also acts as a key factor in modulating the ESC transcriptome. Here in this review, we summarize our current understanding of the transcriptional regulatory network in ESCs, discuss how the control of various signalling pathways could influence pluripotency, and provide a future outlook of ESC research.

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“…A long standing puzzling question has been why most human female ES cell lines carry an Xi, as opposed to female mouse ES cells that always have two active X chromosomes [105][106][107]. Strikingly, in stark contrast to mouse iPS cell lines, currently all available (FGF4-dependent) human female iPS cells do not reactivate the Xi [108], and the ramifications of this Xi retention are proposed to affect studies of X-linked diseases [108].…”

Section: Status Of the X Chromosome Inactivation In Female Cell Repromentioning

confidence: 99%

Reprogramming to pluripotency: stepwise resetting of the epigenetic landscape

2011

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In 2006, the "wall came down" that limited the experimental conversion of differentiated cells into the pluripotent state. In a landmark report, Shinya Yamanaka's group described that a handful of transcription factors (Oct4, Sox2, Klf4 and c-Myc) can convert a differentiated cell back to pluripotency over the course of a few weeks, thus reprograming them into induced pluripotent stem (iPS) cells. The birth of iPS cells started off a rush among researchers to increase the efficiency of the reprogramming process, to reveal the underlying mechanistic events, and allowed the generation of patient-and disease-specific human iPS cells, which have the potential to be converted into relevant specialized cell types for replacement therapies and disease modeling. This review addresses the steps involved in resetting the epigenetic landscape during reprogramming. Apparently, defined events occur during the course of the reprogramming process. Immediately, upon expression of the reprogramming factors, some cells start to divide faster and quickly begin to lose their differentiated cell characteristics with robust downregulation of somatic genes. Only a subset of cells continue to upregulate the embryonic expression program, and finally, pluripotency genes are upregulated establishing an embryonic stem cell-like transcriptome and epigenome with pluripotent capabilities. Understanding reprogramming to pluripotency will inform mechanistic studies of lineage switching, in which differentiated cells from one lineage can be directly reprogrammed into another without going through a pluripotent intermediate.

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Derivation of Pre-X Inactivation Human Embryonic Stem Cells under Physiological Oxygen Concentrations

Cited by 368 publications

References 74 publications

Derivation of naïve human embryonic stem cells

Derivation of naïve human embryonic stem cells

The transcriptional regulation of pluripotency

Reprogramming to pluripotency: stepwise resetting of the epigenetic landscape

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