scholarly journals Quantitative multiplexed ChIP reveals global alterations that shape promoter bivalency in ground state embryonic stem cells

2019 ◽  
Author(s):  
Banushree Kumar ◽  
Simon J Elsässer
Keyword(s):  
Mass Spectrometry ◽  
Stem Cells ◽  
Western Blot ◽  
Ground State ◽  
Embryonic Stem ◽  
Compelling Evidence ◽  
Strong Reduction ◽  
Pluripotent Cells ◽  
Quantitative Western Blot ◽  
Almost All

ABSTRACTTo understand the epigenetic foundation of naïve pluripotent cells, we implement a quantitative multiplexed ChIP-Seq method (MINUTE-ChIP) comparing mouse ESC grown in 2i versus Serum conditions. Combined with quantitative western blot and mass spectrometry, we find compelling evidence for a broad H3K27me3 hypermethylation of the genome, concomitant with the widespread loss of DNA CpG methylation. We show that opposing action of EZH2 and JMJD3/UTX shape the H3K27me3 landscape, with almost all bivalent promoters stably retaining high H3K27me3 levels in 2i. In parallel, we show a mechanistically uncoupled, global decrease of H3K4me3 and strong reduction at bivalent promoters, suggesting that low H3K4me3 and high H3K27me3 levels at bivalent promoters safeguard naïve pluripotency.

A membrane-associated  -catenin/Oct4 complex correlates with ground-state pluripotency in mouse embryonic stem cells

2013 ◽  
Vol 126 (3) ◽  
pp. e1-e1 ◽  
Author(s):  
F. Faunes ◽  
P. Hayward ◽  
S. M. Descalzo ◽  
S. S. Chatterjee ◽  
T. Balayo ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Ground State ◽  
Embryonic Stem ◽  
Mouse Embryonic Stem Cells

scholarly journals Disrupting Interactions Between β-Catenin and Activating TCFs Reconstitutes Ground State Pluripotency in Mouse Embryonic Stem Cells

Stem Cells ◽  
2017 ◽  
Vol 35 (8) ◽  
pp. 1924-1933 ◽  
Author(s):  
Abil Saj ◽  
Sujash S. Chatterjee ◽  
Bowen Zhu ◽  
Engin Cukuroglu ◽  
Tenzin Gocha ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Ground State ◽  
Embryonic Stem ◽  
Mouse Embryonic Stem Cells

scholarly journals Technical Challenges in the Derivation of Human Pluripotent Cells

2011 ◽  
Vol 2011 ◽  
pp. 1-7 ◽  
Author(s):  
Parinya Noisa ◽  
Rangsun Parnpai
Keyword(s):  
Stem Cells ◽  
Cell Biology ◽  
Somatic Cell Nuclear Transfer ◽  
Nuclear Transfer ◽  
Embryonic Stem ◽  
Stem Cell Biology ◽  
Nuclear Reprogramming ◽  
Pluripotent Cells ◽  
Ethical Concerns ◽  
Technical Challenges

It has long been discovered that human pluripotent cells could be isolated from the blastocyst state of embryos and called human embryonic stem cells (ESCs). These cells can be adapted and propagated indefinitely in culture in an undifferentiated manner as well as differentiated into cell representing the three major germ layers: endoderm, mesoderm, and ectoderm. However, the derivation of human pluripotent cells from donated embryos is limited and restricted by ethical concerns. Therefore, various approaches have been explored and proved their success. Human pluripotent cells can also be derived experimentally by the nuclear reprogramming of somatic cells. These techniques include somatic cell nuclear transfer (SCNT), cell fusion and overexpression of pluripotent genes. In this paper, we discuss the technical challenges of these approaches for nuclear reprogramming, involving their advantages and limitations. We will also highlight the possible applications of these techniques in the study of stem cell biology.

scholarly journals Hmga2 is necessary for Otx2-dependent exit of embryonic stem cells from the pluripotent ground state

BMC Biology ◽  
2016 ◽  
Vol 14 (1) ◽  
Author(s):  
Angelica Navarra ◽  
Anna Musto ◽  
Anna Gargiulo ◽  
Giuseppe Petrosino ◽  
Giovanna Maria Pierantoni ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Ground State ◽  
Embryonic Stem

Pluripotent Stem Cells and Reprogrammed Cells in Farm Animals

2011 ◽  
Vol 17 (4) ◽  
pp. 474-497 ◽  
Author(s):  
Monika Nowak-Imialek ◽  
Wilfried Kues ◽  
Joseph W. Carnwath ◽  
Heiner Niemann
Keyword(s):  
Stem Cells ◽  
Pluripotent Stem Cells ◽  
Germ Line ◽  
Embryonic Stem ◽  
Domestic Animals ◽  
Farm Animals ◽  
Pluripotent Cells ◽  
Cell Extracts

AbstractPluripotent cells are unique because of their ability to differentiate into the cell lineages forming the entire organism. True pluripotent stem cells with germ line contribution have been reported for mice and rats. Human pluripotent cells share numerous features of pluripotentiality, but confirmation of their in vivo capacity for germ line contribution is impossible due to ethical and legal restrictions. Progress toward derivation of embryonic stem cells from domestic species has been made, but the derived cells were not able to produce germ line chimeras and thus are termed embryonic stem-like cells. However, domestic animals, in particular the domestic pig (Sus scrofa), are excellent large animals models, in which the clinical potential of stem cell therapies can be studied. Reprogramming technologies for somatic cells, including somatic cell nuclear transfer, cell fusion, in vitro culture in the presence of cell extracts, in vitro conversion of adult unipotent spermatogonial stem cells into germ line derived pluripotent stem cells, and transduction with reprogramming factors have been developed with the goal of obtaining pluripotent, germ line competent stem cells from domestic animals. This review summarizes the present state of the art in the derivation and maintenance of pluripotent stem cells in domestic animals.

scholarly journals Critical role for P53 in regulating the cell cycle of ground state embryonic stem cells

2019 ◽  
Author(s):  
Menno ter Huurne ◽  
Tianran Peng ◽  
Guoqiang Yi ◽  
Guido van Mierlo ◽  
Hendrik Marks ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Ground State ◽  
Critical Role ◽  
Embryonic Stem ◽  
Phase Control ◽  
G1 Phase ◽  
Rna Seq ◽  
Rb Protein ◽  
Uncontrolled Cell Proliferation

AbstractMouse Embryonic Stem Cells (ESCs) grown in serum-supplemented conditions are characterized by an extremely short G1-phase due to the lack of G1-phase control. Concordantly, the G1-phase-specific P53-P21 pathway is compromised in serum ESCs. Here we provide evidence that P53 is activated upon transition of serum ESCs to their pluripotent ground state using serum-free 2i conditions and modulates G1-phase progression. Our data shows that the elongated G1-phase characteristic of ground state ESCs is dependent on P53. RNA-seq and ChIP-seq analyses reveal that P53 directly regulates the expression of the Retinoblastoma (RB) protein and that the hypo-phosphorylated, active RB protein plays a key role in G1-phase control. Our findings suggest that the P53-P21 pathway is active in ground state 2i ESCs and that its role in the G1-checkpoint is abolished in serum ESCs. Taken together, the data reveals a mechanism by which inactivation of P53 can lead to loss of RB and uncontrolled cell proliferation.

scholarly journals Identification of Novel miRNAs in Mouse Embryonic Stem Cells, Embryonic Fibroblasts, and Reprogrammed Pluripotent Cells

2019 ◽  
Author(s):  
Botao Zhao ◽  
Chunsun Fan
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Embryonic Stem Cells ◽  
De Novo ◽  
Embryonic Stem ◽  
Induced Pluripotent Stem Cell ◽  
Mouse Embryonic Stem Cells ◽  
Pluripotent Cells ◽  
Embryonic Fibroblasts ◽  
Species Specific

AbstractMicroRNAs (miRNAs) are a class of non-coding small RNAs that function in almost every known cellular activity. MiRNAs play an important role in gene regulation that controls embryonic stem cell (ESC) pluripotency and differentiation, as well as induced pluripotent stem cell (iPSC) reprogramming. In this study, we identified nine novel miRNAs by mining the deep sequencing dataset from mouse embryonic stem cells, mouse embryonic fibroblasts (MEF) and three kinds of reprogrammed pluripotent cells. Most of them are non-conserved but species-specific and cell-specific miRNAs. Eight miRNAs are derived from gene introns, including a “mirtron” miRNA, miR-novel-41. We also showed that miR-novel-27 is a mouse-specific miRNA and the 5′ arm of its precursor hairpin, embedding the mature miR-novel-27, uniquely exists in mouse species but not in any other Placentalia animals. Notably, the 5′ arm of the pre-miR-novel-27 hairpin shows nearly perfect palindrome to the 3′ arm suggesting that it was generated by inverted duplication of the 3′ arm. By this mechanism, the pre-miR-novel-27 hairpin was de novo gained in the mouse genome. This is a new type of de novo miRNA emergence mechanism in animals, which we called “inverted local half hairpin duplication” here. In addition, very limited nucleotide mutants accumulated on the newly emerged 5′ arm since its birth suggesting an especially young evolutionary history of the miR-novel-27 gene.

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