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Alessandro Fiorenzano, Emilia Pascale, Cristina D'Aniello, Dario Acampora, Cecilia Bassalert, Francesco Russo, Gennaro Andolfi, Mauro Biffoni, Federica Francescangeli, Ann Zeuner, Claudia Angelini, Claire Chazaud, Eduardo J Patriarca, Annalisa Fico, Gabriella Minchiotti
Known molecular determinants of developmental plasticity are mainly transcription factors, while the extrinsic regulation of this process has been largely unexplored. Here we identify Cripto as one of the earliest epiblast markers and a key extracellular determinant of the naive and primed pluripotent states. We demonstrate that Cripto sustains mouse embryonic stem cell (ESC) self-renewal by modulating Wnt/β-catenin, whereas it maintains mouse epiblast stem cell (EpiSC) and human ESC pluripotency through Nodal/Smad2...
2016: Nature Communications
Tara TeSlaa, Andrea C Chaikovsky, Inna Lipchina, Sandra L Escobar, Konrad Hochedlinger, Jing Huang, Thomas G Graeber, Daniel Braas, Michael A Teitell
Pluripotent stem cells (PSCs) can self-renew or differentiate from naive or more differentiated, primed, pluripotent states established by specific culture conditions. Increased intracellular α-ketoglutarate (αKG) was shown to favor self-renewal in naive mouse embryonic stem cells (mESCs). The effect of αKG or αKG/succinate levels on differentiation from primed human PSCs (hPSCs) or mouse epiblast stem cells (EpiSCs) remains unknown. We examined primed hPSCs and EpiSCs and show that increased αKG or αKG-to-succinate ratios accelerate, and elevated succinate levels delay, primed PSC differentiation...
September 13, 2016: Cell Metabolism
Dario Acampora, Daniela Omodei, Giuseppe Petrosino, Arcomaria Garofalo, Marco Savarese, Vincenzo Nigro, Luca Giovanni Di Giovannantonio, Vincenzo Mercadante, Antonio Simeone
Mouse embryonic stem cells (ESCs) and the inner cell mass (ICM)-derived epiblast exhibit naive pluripotency. ESC-derived epiblast stem cells (EpiSCs) and the postimplantation epiblast exhibit primed pluripotency. Although core pluripotency factors are well-characterized, additional regulators, including Otx2, recently have been shown to function during the transition from naive to primed pluripotency. Here we uncover a role for Otx2 in the control of the naive pluripotent state. We analyzed Otx2-binding activity in ESCs and EpiSCs and identified Nanog, Oct4, and Sox2 as direct targets...
June 21, 2016: Cell Reports
Lu Song, Jun Chen, Guangdun Peng, Ke Tang, Naihe Jing
Mouse pluripotent cells, such as embryonic stem cells (ESCs) and epiblast stem cells (EpiSCs), provide excellent in vitro systems to study imperative pre- and postimplantation events of in vivo mammalian development. It is known that mouse ESCs are dynamic heterogeneous populations. However, it remains largely unclear whether and how EpiSCs possess heterogeneity and plasticity similar to that of ESCs. Here, we show that EpiSCs are discriminated by the expression of a specific marker T (Brachyury) into two populations...
July 15, 2016: Journal of Biological Chemistry
Damir Jacob Illich, Miao Zhang, Andrei Ursu, Rodrigo Osorno, Kee-Pyo Kim, Juyong Yoon, Marcos J Araúzo-Bravo, Guangming Wu, Daniel Esch, Davood Sabour, Douglas Colby, Kathrin S Grassme, Jiayu Chen, Boris Greber, Susanne Höing, Wiebke Herzog, Slava Ziegler, Ian Chambers, Shaorong Gao, Herbert Waldmann, Hans R Schöler
It has previously been reported that mouse epiblast stem cell (EpiSC) lines comprise heterogeneous cell populations that are functionally equivalent to cells of either early- or late-stage postimplantation development. So far, the establishment of the embryonic stem cell (ESC) pluripotency gene regulatory network through the widely known chemical inhibition of MEK and GSK3beta has been impractical in late-stage EpiSCs. Here, we show that chemical inhibition of casein kinase 1alpha (CK1alpha) induces the conversion of recalcitrant late-stage EpiSCs into ESC pluripotency...
April 14, 2016: Cell Reports
Li Ding, Maciej Paszkowski-Rogacz, Maria Winzi, Debojyoti Chakraborty, Mirko Theis, Sukhdeep Singh, Giovanni Ciotta, Ina Poser, Assen Roguev, Wai Kit Chu, Chunaram Choudhary, Matthias Mann, A Francis Stewart, Nevan Krogan, Frank Buchholz
We combine a genome-scale RNAi screen in mouse epiblast stem cells (EpiSCs) with genetic interaction, protein localization, and "protein-level dependency" studies-a systematic technique that uncovers post-transcriptional regulation-to delineate the network of factors that control the expression of Oct4, a key regulator of pluripotency. Our data signify that there are similarities, but also fundamental differences in Oct4 regulation in EpiSCs versus embryonic stem cells (ESCs). Through multiparametric data analyses, we predict that Tox4 is associating with the Paf1C complex, which maintains cell identity in both cell types, and validate that this protein-protein interaction exists in ESCs and EpiSCs...
August 26, 2015: Cell Systems
Andrei Ursu, Damir J Illich, Yasushi Takemoto, Arthur T Porfetye, Miao Zhang, Andreas Brockmeyer, Petra Janning, Nobumoto Watanabe, Hiroyuki Osada, Ingrid R Vetter, Slava Ziegler, Hans R Schöler, Herbert Waldmann
The discovery of novel small molecules that induce stem cell reprogramming and give efficient access to pluripotent stem cells is of major importance for potential therapeutic applications and may reveal novel insights into the factors controlling pluripotency. Chemical reprogramming of mouse epiblast stem cells (EpiSCs) into cells corresponding to embryonic stem cells (cESCs) is an inefficient process. In order to identify small molecules that promote this cellular transition, we analyzed the LOPAC library in a phenotypic screen monitoring Oct4-GFP expression and identified triamterene (TR) as initial hit...
April 21, 2016: Cell Chemical Biology
Smita Sudheer, Jinhua Liu, Matthias Marks, Frederic Koch, Anna Anurin, Manuela Scholze, Anna Dorothea Senft, Lars Wittler, Karol Macura, Phillip Grote, Bernhard G Herrmann
Presomitic mesoderm (PSM) cells are the precursors of the somites, which flank both sides of the neural tube and give rise to the musculo-skeletal system shaping the vertebrate body. WNT and FGF signaling control the formation of both the PSM and the somites and show a graded distribution with highest levels in the posterior PSM. We have used reporters for the mesoderm/PSM control genes T, Tbx6, and Msgn1 to investigate the differentiation of mouse ESCs from the naïve state via EpiSCs to PSM cells. Here we show that the activation of WNT signaling by CHIR99021 (CH) in combination with FGF ligand induces embryo-like PSM at high efficiency...
July 2016: Stem Cells
Hui Zhang, Srimonta Gayen, Jie Xiong, Bo Zhou, Avinash K Shanmugam, Yuqing Sun, Hacer Karatas, Liu Liu, Rajesh C Rao, Shaomeng Wang, Alexey I Nesvizhskii, Sundeep Kalantry, Yali Dou
The interconversion between naive and primed pluripotent states is accompanied by drastic epigenetic rearrangements. However, it is unclear whether intrinsic epigenetic events can drive reprogramming to naive pluripotency or if distinct chromatin states are instead simply a reflection of discrete pluripotent states. Here, we show that blocking histone H3K4 methyltransferase MLL1 activity with the small-molecule inhibitor MM-401 reprograms mouse epiblast stem cells (EpiSCs) to naive pluripotency. This reversion is highly efficient and synchronized, with more than 50% of treated EpiSCs exhibiting features of naive embryonic stem cells (ESCs) within 3 days...
April 7, 2016: Cell Stem Cell
Pierre Osteil, Joshua Studdert, Emilie Wilkie, Nicolas Fossat, Patrick P L Tam
Conventionally, mouse epiblast stem cells (EpiSCs) are derived directly from the epiblast or ectoderm germ layer of the post-implantation embryo. Self-renewing and multipotent EpiSC-like stem cells can also be derived by the conversion of embryonic stem cells (ESCs) via the provision of culture conditions that enable the maintenance of the EpiSCs. Here, we outline an experimental procedure for deriving EpiSCs from post-implantation chimeric embryos that are generated using genome-edited ESCs. This strategy enables the production of EpiSCs where (i) no genetically modified animals or ESCs are available, (ii) the impact of the genetic modification on post-implantation development, which may influence the property of the EpiSCs, is requisite knowledge for using the EpiSC for a specific investigation, and (iii) multiple editing of the genome is desirable to modify the biological attributes of the EpiSCs for studying, for example, the gene network activity on the trajectory of lineage differentiation and tissue morphogenesis...
April 2016: Differentiation; Research in Biological Diversity
Matteo Moretto Zita, Francesca Soncin, David Natale, Donald Pizzo, Mana Parast
Appropriate self-renewal and differentiation of trophoblast stem cells (TSCs) are key factors for proper placental development and function and, in turn, for appropriate in utero fetal growth. To identify novel TSC-specific genes, we performed genome-wide expression profiling of TSCs, embryonic stem cells, epiblast stem cells, and mouse embryo fibroblasts, derived from mice of the same genetic background. Our analysis revealed a high expression of Sox21 in TSCs compared with other cell types. Sox21 levels were high in undifferentiated TSCs and were dramatically reduced upon differentiation...
December 11, 2015: Journal of Biological Chemistry
Maria Fernanda Forni, Aline Ramos Maia Lobba, Alexandre Hamilton Pereira Ferreira, Mari Cleide Sogayar
The skin is a rich source of readily accessible stem cells. The level of plasticity afforded by these cells is becoming increasingly important as the potential of stem cells in Cell Therapy and Regenerative Medicine continues to be explored. Several protocols described single type stem cell isolation from skin; however, none of them afforded simultaneous isolation of more than one population. Herein, we describe the simultaneous isolation and characterization of three stem cell populations from the dermis and epidermis of murine skin, namely Epidermal Stem Cells (EpiSCs), Skin-derived Precursors (SKPs) and Mesenchymal Stem Cells (MSCs)...
2015: PloS One
Constantinos Economou, Anestis Tsakiridis, Filip J Wymeersch, Sabrina Gordon-Keylock, Robert E Dewhurst, Dawn Fisher, Alexander Medvinsky, Andrew J H Smith, Valerie Wilson
BACKGROUND: Pluripotent cells are present in early embryos until the levels of the pluripotency regulator Oct4 drop at the beginning of somitogenesis. Elevating Oct4 levels in explanted post-pluripotent cells in vitro restores their pluripotency. Cultured pluripotent cells can participate in normal development when introduced into host embryos up to the end of gastrulation. In contrast, pluripotent cells efficiently seed malignant teratocarcinomas in adult animals. In humans, extragonadal teratomas and teratocarcinomas are most frequently found in the sacrococcygeal region of neonates, suggesting that these tumours originate from cells in the posterior of the embryo that either reactivate or fail to switch off their pluripotent status...
October 9, 2015: BMC Developmental Biology
Siu-Shan Mak, Cantas Alev, Hiroki Nagai, Anna Wrabel, Yoko Matsuoka, Akira Honda, Guojun Sheng, Raj K Ladher
Innate pluripotency of mouse embryos transits from naive to primed state as the inner cell mass differentiates into epiblast. In vitro, their counterparts are embryonic (ESCs) and epiblast stem cells (EpiSCs), respectively. Activation of the FGF signaling cascade results in mouse ESCs differentiating into mEpiSCs, indicative of its requirement in the shift between these states. However, only mouse ESCs correspond to the naive state; ESCs from other mammals and from chick show primed state characteristics. Thus, the significance of the naive state is unclear...
2015: ELife
Masaki Kinoshita
In mice, three pluripotent stem cell lines have been established from different stage of developing embryo, which are embryonic stem (ES) cell, post-implantation epiblast stem cell (EpiSC), and embryonic germ (EG) cell. ES cell and EG cell share many common features including factor requirement, colony morphology, and gene expression pattern. On the other hand, EpiSC needs different external signal inputs, exhibits flattened colony morphology, and a different set of gene expression patterns. In addition, the germ line competency of EpiSCs is still unclear...
2015: Reproductive Medicine and Biology
Zofia Eliza Madeja, Kamila Hryniewicz, Maciej Orsztynowicz, Piotr Pawlak, Anna Perkowska
Despite many attempts, true bovine embryonic stem cells (bESC) still remain elusive. The WNT pathway has been associated with stem cell control in vertebrates and its role in pluripotency maintenance has been proven for several mammalian species, including rodents and primates. Thus, we have aimed to investigate the effect of WNT activation on pluripotency marker gene expression in the inner cell mass (ICM) and the trophectoderm (TE) and to study the derivation potential of primary bESC lines from blastocysts obtained in the presence of the glycogen synthase kinase 3 inhibitor (GSK3i)...
October 15, 2015: Stem Cells and Development
Uri Weissbein, Nissim Benvenisty
Pluripotent stem cells capture the imagination since they can differentiate into all cell types in our body. Recent evidence suggests that in addition to embryonic stem cells (ESCs) and epiblast stem cells (EpiSCs), a new type of region-selective pluripotent stem cells (rsPSCs) exists, possessing unique spatial and molecular characteristics.
August 2015: Cell Research
Nika Shakiba, Carl A White, Yonatan Y Lipsitz, Ayako Yachie-Kinoshita, Peter D Tonge, Samer M I Hussein, Mira C Puri, Judith Elbaz, James Morrissey-Scoot, Mira Li, Javier Munoz, Marco Benevento, Ian M Rogers, Jacob H Hanna, Albert J R Heck, Bernd Wollscheid, Andras Nagy, Peter W Zandstra
Reprogramming is a dynamic process that can result in multiple pluripotent cell types emerging from divergent paths. Cell surface protein expression is a particularly desirable tool to categorize reprogramming and pluripotency as it enables robust quantification and enrichment of live cells. Here we use cell surface proteomics to interrogate mouse cell reprogramming dynamics and discover CD24 as a marker that tracks the emergence of reprogramming-responsive cells, while enabling the analysis and enrichment of transgene-dependent (F-class) and -independent (traditional) induced pluripotent stem cells (iPSCs) at later stages...
2015: Nature Communications
Lingyu Li, Lu Song, Chang Liu, Jun Chen, Guangdun Peng, Ran Wang, Pingyu Liu, Ke Tang, Janet Rossant, Naihe Jing
The ectoderm has the capability to generate epidermis and neuroectoderm and plays imperative roles during the early embryonic development. Our recent study uncovered a region with ectodermal progenitor potential in mouse embryo at embryonic day 7.0 and revealed that Nodal inhibition is essential for its formation. Here, we demonstrate that through brief inhibition of Nodal signaling in vitro, mouse embryonic stem cell (ESC)-derived epiblast stem cells (ESD-EpiSCs) could be committed to transient ectodermal progenitor populations, which possess the ability to give rise to neural or epidermal ectoderm in the absence or presence of BMP4, respectively...
October 2015: Journal of Molecular Cell Biology
Simone Aparecida Siqueira Fonseca, Roberta Montero Costas, Lygia Veiga Pereira
Normal mouse pluripotent stem cells were originally derived from the inner cell mass (ICM) of blastocysts and shown to be the in vitro equivalent of those pre-implantation embryonic cells, and thus were called embryonic stem cells (ESCs). More than a decade later, pluripotent cells were isolated from the ICM of human blastocysts. Despite being called human ESCs, these cells differ significantly from mouse ESCs, including different morphology and mechanisms of control of pluripotency, suggesting distinct embryonic origins of ESCs from the two species...
April 26, 2015: World Journal of Stem Cells
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