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Jiangwa Xing, Yue Cao, Yang Yu, Huan Li, Ziwei Song, Hanry Yu
Exposure to teratogenic chemicals during pregnancy may cause severe birth defects. Due to high inter-species variation of drug responses as well as financial and ethical burdens, despite the widely use of in vivo animal tests, it's crucial to develop highly predictive human pluripotent stem cell (hPSC)-based in vitro assays to identify potential teratogens. Previously we have shown that the morphological disruption of mesoendoderm patterns formed by geometrically-confined cell differentiation and migration using hPSCs could potentially serve as a sensitive morphological marker in teratogen detection...
August 17, 2017: Scientific Reports
Yang Li, Patrick M Brauer, Jastaranpreet Singh, Sintia Xhiku, Kogulan Yoganathan, Juan Carlos Zúñiga-Pflücker, Michele K Anderson
Hematopoietic stem cells arise from mesoderm-derived hemogenic endothelium (HE) during embryogenesis in a process termed endothelial-hematopoietic transition (EHT). To better understand the gene networks that control this process, we investigated the role of the transcription factor HEB (TCF12) by disrupting the TCF12 gene locus in human embryonic stem cells (hESCs) and inducing them to differentiate toward hematopoietic outcomes. HEB-deficient hESCs retained key features of pluripotency, including expression of SOX2 and SSEA-4 and teratoma formation, while NANOG expression was reduced...
September 12, 2017: Stem Cell Reports
Tomohiko Akiyama, Shunichi Wakabayashi, Atsumi Soma, Saeko Sato, Yuhki Nakatake, Mayumi Oda, Miyako Murakami, Miki Sakota, Nana Chikazawa-Nohtomi, Shigeru B H Ko, Minoru S H Ko
Harnessing epigenetic regulation is crucial for the efficient and proper differentiation of pluripotent stem cells (PSCs) into desired cell types. Histone H3 lysine 27 trimethylation (H3K27me3) functions as a barrier against cell differentiation through the suppression of developmental gene expression in PSCs. Here, we have generated human PSC (hPSC) lines in which genome-wide reduction of H3K27me3 can be induced by ectopic expression of the catalytic domain of the histone demethylase JMJD3 (called JMJD3c)...
October 15, 2016: Development
Geetika Sahni, Jun Yuan, Yi-Chin Toh
Human pluripotent stem cells (hPSCs), including embryonic stem cells and induced pluripotent stem cells, have the intrinsic ability to differentiate into all three germ layers. This makes them an attractive cell source for regenerative medicine and experimental modeling of normal and diseased organogenesis. However, the differentiation of hPSCs in vitro is heterogeneous and spatially disordered. Cell micropatterning technologies potentially offer the means to spatially control stem cell microenvironments and organize the resultant differentiation fates...
June 17, 2016: Journal of Visualized Experiments: JoVE
Tatsuki Nagasawa, Mari Kawaguchi, Tohru Yano, Kaori Sano, Masataka Okabe, Shigeki Yasumasu
Hatching gland cells (HGCs) originate from different germ layers between frogs and teleosts, although the hatching enzyme genes are orthologous. Teleostei HGCs differentiate in the mesoendodermal cells at the anterior end of the involved hypoblast layer (known as the polster) in late gastrula embryos. Conversely, frog HGCs differentiate in the epidermal cells at the neural plate border in early neurula embryos. To infer the transition in the developmental origin of HGCs, we studied two basal ray-finned fishes, bichir (Polypterus) and sturgeon...
June 2016: Zoological Science
Jiangwa Xing, Yi-Chin Toh, Shuoyu Xu, Hanry Yu
Unintended exposure to teratogenic compounds can lead to various birth defects; however current animal-based testing is limited by time, cost and high inter-species variability. Here, we developed a human-relevant in vitro model, which recapitulated two cellular events characteristic of embryogenesis, to identify potentially teratogenic compounds. We spatially directed mesoendoderm differentiation, epithelial-mesenchymal transition and the ensuing cell migration in micropatterned human pluripotent stem cell (hPSC) colonies to collectively form an annular mesoendoderm pattern...
2015: Scientific Reports
M Duque, M N Biancardi, J H Galiguis, C E Pope, C Dumas, G Wang, M C Gómez
Different feeder cells (FC) influence the isolation, proliferation, and self-renewal of cat embryonic stem cells (cat ESC; Gómez et al. 2010 Theriogenology 74, 498-515) possibly by secretion of growth factors that affect intracellular signalling pathways involved in self-renewal. Supplementation of the culture medium with fibroblast growth factor (FGF) stimulates the secretion of Activin A in mouse and human FC, which enhances undifferentiation in human ESC (Eiselleova et al. 2008 Int. J. Dev. Biol. 52, 353-363)...
December 2014: Reproduction, Fertility, and Development
Sumit Bhattacharyya, Leo Feferman, Joanne K Tobacman
In cultured human colonic epithelial cells and mouse colonic tissue, exposure to the common food additive carrageenan leads to inflammation, activation of Wnt signaling, increased Wnt9A expression, and decline in the activity of the enzyme arylsulfatase B (ARSB; N-acetylgalactosamine-4-sulfatase). In this study, the novel transcriptional mechanism by which carrageenan and decline in ARSB increase Wnt9A expression in NCM460 and HT-29 human colonic epithelial cells and in mouse colon is presented. Increased expression of Wnt9A has been associated with multiple malignancies, including colon carcinoma, and with ectodermal and mesoendodermal morphogenesis...
June 20, 2014: Journal of Biological Chemistry
Oluwaseun Akeju, Brandi N Davis-Dusenbery, Seth H Cassel, Justin K Ichida, Kevin Eggan
Preclinical and clinical evidence implicates N-methyl-d-aspartate receptor (NMDAr) signaling in early embryological development. However, the role of NMDAr signaling in early development has not been well studied. Here, we use a mouse embryonic stem cell model to perform a step-wise exploration of the effects of NMDAr signaling on early cell fate specification. We found that antagonism of the NMDAr impaired specification into the neuroectodermal and mesoendodermal cell lineages, with little or no effect on specification of the extraembryonic endoderm cell lineage...
May 2014: Neurotoxicology and Teratology
Ikuko Yazaki, Toko Tsurugaya, Luigia Santella, Jong Tai Chun, Gabriele Amore, Shinichiro Kusunoki, Akiko Asada, Tatsuru Togo, Koji Akasaka
Sea urchin embryos initiate cell specifications at the 16-cell stage by forming the mesomeres, macromeres and micromeres according to the relative position of the cells in the animal-vegetal axis. The most vegetal cells, micromeres, autonomously differentiate into skeletons and induce the neighbouring macromere cells to become mesoendoderm in the β-catenin-dependent Wnt8 signalling pathway. Although the underlying molecular mechanism for this progression is largely unknown, we have previously reported that the initial events might be triggered by the Ca2+ influxes through the egg-originated L-type Ca2+ channels distributed asymmetrically along the animal-vegetal axis and through the stretch-dependent Ca2+channels expressed specifically in the micromere at the 4th cleavage...
June 2015: Zygote: the Biology of Gametes and Early Embryos
Percival Sangel, Masahiro Oka, Yoshihiro Yoneda
Members of the Importin-β family recognize nuclear localization signals (NLS) and nuclear export signals (NES). These proteins play important roles in various nucleocytoplasmic transport processes in cells. Here, we examined the expression patterns of 21 identified Importin-β genes in mouse embryonic stem cells (mESCs), mouse embryonic fibroblast (MEF) and mESCs differentiated into neural ectoderm (NE) or mesoendoderm (ME). We observed striking differences in the Importin-β mRNA expression levels within these cell types...
2014: FEBS Open Bio
Joshua W Ogony, Evangelia Malahias, Rajanikanth Vadigepalli, Helen Anni
The transcription factors Sox2, Oct4, and Nanog regulate within a narrow dose-range embryonic stem (ES) cell pluripotency and cell lineage commitment. Excess of Oct4 relative to Sox2 guides cells to mesoendoderm (ME), while abundance of Sox2 promotes neuroectoderm (NE) formation. Literature does not address whether ethanol interferes with these regulatory interactions during neural development. We hypothesized that ethanol exposure of ES cells in early differentiation causes an imbalance of Oct4 and Sox2 that diverts cells away from NE to ME lineage, consistent with the teratogenesis effects caused by prenatal alcohol exposure...
August 1, 2013: Stem Cells and Development
Harsha Pawani, Punam Nagvenkar, Prasad Pethe, Deepa Bhartiya
Human embryonic stem cells (hESCs) have the ability to differentiate into all the three lineages and are an ideal starting material to obtain cells of desired lineage for regenerative medicine. Continued efforts are needed to evolve more robust protocols to obtain cells of desired lineages and in larger numbers. Also, it has now been realized that rather than transplanting fully committed cells differentiated in vitro, it may be ideal to transplant committed progenitors which retain the intrinsic ability to proliferate and also differentiate better into multiple lineages based on the in vivo cues...
January 2013: In Vitro Cellular & Developmental Biology. Animal
Yujun Zhang, Bingyu Mao
The secreted Wnt signaling inhibitor Dickkopf1 (Dkk1) plays key role in vertebrate head induction. Its receptor Kremen synergizes with Dkk1 in Wnt inhibition. Here we have carried out expression and functional studies of the Dkk and Kremen genes in amphioxus (Branchiostoma belcheri). During embryonic and larval development, BbDkk1/2/4 is expressed in the posterior mesoendoderm, anterior somatic mesoderm and the pharyngeal regions. Its expression becomes restricted to the pharyngeal region on the left side at larval stages...
September 2010: Journal of Genetics and Genomics, Yi Chuan Xue Bao
Veronica Ramos-Mejia, Gustavo J Melen, Laura Sanchez, Ivan Gutierrez-Aranda, Gertrudis Ligero, Jose L Cortes, Pedro J Real, Clara Bueno, Pablo Menendez
Lineage-specific differentiation potential varies among different human pluripotent stem cell (hPSC) lines, becoming therefore highly desirable to prospectively know which hPSC lines exhibit the highest differentiation potential for a certain lineage. We have compared the hematopoietic potential of 14 human embryonic stem cell (hESC)/induced pluripotent stem cell (iPSC) lines. The emergence of hemogenic progenitors, primitive and mature blood cells, and colony-forming unit (CFU) potential was analyzed at different time points...
December 2010: Molecular Therapy: the Journal of the American Society of Gene Therapy
J Liang, Y-J Wang, Y Tang, N Cao, J Wang, H-T Yang
Ca(2+) signals generated by inositol 1,4,5-trisphosphate receptors (IP(3)Rs) are crucial for cellular processes such as apoptosis and differentiation. However, the exact roles of IP(3)Rs and their contributions to Ca(2+) signals in pluripotent embryonic stem (ES) cell behaviors remain largely unknown. In this study, we showed that the expression of type 3 IP(3)R (IP(3)R3) was transiently downregulated with a concomitant increase in apoptosis at the early differentiation stage of murine ES cells. Knockdown of IP(3)R3 by small interfering RNA increased apoptosis in differentiating cells but not in undifferentiated ES cells...
July 2010: Cell Death and Differentiation
Sheng Gao, Claudio Alarcón, Gopal Sapkota, Sadia Rahman, Pan-Yu Chen, Nina Goerner, Maria J Macias, Hediye Erdjument-Bromage, Paul Tempst, Joan Massagué
TGF-beta induces phosphorylation of the transcription factors Smad2 and Smad3 at the C terminus as well as at an interdomain linker region. TGF-beta-induced linker phosphorylation marks the activated Smad proteins for proteasome-mediated destruction. Here, we identify Nedd4L as the ubiquitin ligase responsible for this step. Through its WW domain, Nedd4L specifically recognizes a TGF-beta-induced phosphoThr-ProTyr motif in the linker region, resulting in Smad2/3 polyubiquitination and degradation. Nedd4L is not interchangeable with Smurf1, a ubiquitin ligase that targets BMP-activated, linker-phosphorylated Smad1...
November 13, 2009: Molecular Cell
Sakie Hosoya-Ohmura, Naomi Mochizuki, Mikiko Suzuki, Osamu Ohneda, Kinuko Ohneda, Masayuki Yamamoto
Vertebrate GATA transcription factors have been classified into two subgroups; GATA-1, GATA-2, and GATA-3 are expressed in hematopoietic cells, whereas GATA-4, GATA-5, and GATA-6 are expressed in mesoendoderm-derived tissues. We previously discovered that expression of GATA-2 or GATA-3 under the transcriptional control for the Gata1 gene eliminates lethal anemia in Gata1 germ line mutant mice (Gata1.05/Y). Here, we show that the GATA-4 expression by the same regulatory cassette prolongs the life span of Gata1...
October 27, 2006: Journal of Biological Chemistry
Tetsuya Koide, Tadayoshi Hayata, Ken W Y Cho
Development is controlled by a complex series of events requiring sequential gene activation. Understanding the logic of gene networks during development is necessary for a complete understanding of how genes contribute to phenotype. Pioneering work initiated in the sea urchin and Drosophila has demonstrated that reasonable transcriptional regulatory network diagrams representing early development in multicellular animals can be generated through use of appropriate genomic, genetic, and biochemical tools. Establishment of similar regulatory network diagrams for vertebrate development is a necessary step...
April 5, 2005: Proceedings of the National Academy of Sciences of the United States of America
Chi Zhang, Tamara Basta, Shana R Fawcett, M W Klymkowsky
In zebrafish, the divergent F-type SOX casanova acts downstream of Nodal signaling to specify endoderm. While no casanova orthologs have been identified in tetrapods, the F-type SOX, SOX7, is supplied maternally in Xenopus (Fawcett and Klymkowsky, 2004. GER 4, 29). Subsequent RT-PCR and section-based in situ hybridization analyses indicate that SOX7 mRNA is localized to the vegetal region of the blastula-stage embryo. Overexpression and maternal depletion studies reveal that the T-box transcription factor VegT, which initiates mesoendodermal differentiation, directly regulates SOX7 expression...
February 15, 2005: Developmental Biology
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