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https://www.readbyqxmd.com/read/27918544/a-balance-between-elongation-and-trimming-regulates-telomere-stability-in-stem-cells
#1
Teresa Rivera, Candy Haggblom, Sandro Cosconati, Jan Karlseder
Telomere length maintenance ensures self-renewal of human embryonic stem cells (hESCs) and induced pluripotent stem cells (hiPSCs); however, the mechanisms governing telomere length homeostasis in these cell types are unclear. Here, we report that telomere length is determined by the balance between telomere elongation, which is mediated by telomerase, and telomere trimming, which is controlled by XRCC3 and Nbs1, homologous recombination proteins that generate single-stranded C-rich telomeric DNA and double-stranded telomeric circular DNA (T-circles), respectively...
December 5, 2016: Nature Structural & Molecular Biology
https://www.readbyqxmd.com/read/27917311/rock-inhibition-promotes-attachment-proliferation-and-wound-closure-in-human-embryonic-stem-cell-derived-retinal-pigmented-epithelium
#2
Roxanne H Croze, William J Thi, Dennis O Clegg
PURPOSE: Nonexudative (dry) age-related macular degeneration (AMD), a leading cause of blindness in the elderly, is associated with the loss of retinal pigmented epithelium (RPE) cells and the development of geographic atrophy, which are areas devoid of RPE cells and photoreceptors. One possible treatment option would be to stimulate RPE attachment and proliferation to replace dying/dysfunctional RPE and bring about wound repair. Clinical trials are underway testing injections of RPE cells derived from pluripotent stem cells to determine their safety and efficacy in treating AMD...
November 2016: Translational Vision Science & Technology
https://www.readbyqxmd.com/read/27912097/lncpress1-is-a-p53-regulated-lncrna-that-safeguards-pluripotency-by-disrupting-sirt6-mediated-de-acetylation-of-histone-h3k56
#3
Abhinav K Jain, Yuanxin Xi, Ryan McCarthy, Kendra Allton, Kadir C Akdemir, Lalit R Patel, Bruce Aronow, Chunru Lin, Wei Li, Liuqing Yang, Michelle C Barton
Recent evidence suggests that lncRNAs play an integral regulatory role in numerous functions, including determination of cellular identity. We determined global expression (RNA-seq) and genome-wide profiles (ChIP-seq) of histone post-translational modifications and p53 binding in human embryonic stem cells (hESCs) undergoing differentiation to define a high-confidence set of 40 lncRNAs, which are p53 transcriptional targets. We focused on lncRNAs highly expressed in pluripotent hESCs and repressed by p53 during differentiation to identify lncPRESS1 as a p53-regulated transcript that maintains hESC pluripotency in concert with core pluripotency factors...
December 1, 2016: Molecular Cell
https://www.readbyqxmd.com/read/27909687/morphogenesis-of-human-embryonic-stem-cells-into-mature-neurons-under-in-vitro-culture-conditions
#4
Geeta Shroff
AIM: To describe the morphogenesis of different neuronal cells from the human embryonic stem cell (hESC) line, SCT-N, under in vitro culture conditions. METHODS: The directed neuronal cell line was produced from a single, spare, pre-implantation stage fertilized ovum that was obtained during a natural in vitro fertilization process. The hESCs were cultured and maintained as per our proprietary in-house technology in a Good Manufacturing Practice, Good Laboratory Practice and Good Tissue Practice compliant laboratory...
November 20, 2016: World Journal of Experimental Medicine
https://www.readbyqxmd.com/read/27899664/integration-defective-lentiviral-vector-mediates-efficient-gene-editing-through-homology-directed-repair-in-human-embryonic-stem-cells
#5
Yebo Wang, Yingjia Wang, Tammy Chang, He Huang, Jiing-Kuan Yee
Human embryonic stem cells (hESCs) are used as platforms for disease study, drug screening and cell-based therapy. To facilitate these applications, it is frequently necessary to genetically manipulate the hESC genome. Gene editing with engineered nucleases enables site-specific genetic modification of the human genome through homology-directed repair (HDR). However, the frequency of HDR remains low in hESCs. We combined efficient expression of engineered nucleases and integration-defective lentiviral vector (IDLV) transduction for donor template delivery to mediate HDR in hESC line WA09...
November 28, 2016: Nucleic Acids Research
https://www.readbyqxmd.com/read/27892468/somatic-increase-of-cct8-mimics-proteostasis-of-human-pluripotent-stem-cells-and-extends-c-elegans-lifespan
#6
Alireza Noormohammadi, Amirabbas Khodakarami, Ricardo Gutierrez-Garcia, Hyun Ju Lee, Seda Koyuncu, Tim König, Christina Schindler, Isabel Saez, Azra Fatima, Christoph Dieterich, David Vilchez
Human embryonic stem cells can replicate indefinitely while maintaining their undifferentiated state and, therefore, are immortal in culture. This capacity may demand avoidance of any imbalance in protein homeostasis (proteostasis) that would otherwise compromise stem cell identity. Here we show that human pluripotent stem cells exhibit enhanced assembly of the TRiC/CCT complex, a chaperonin that facilitates the folding of 10% of the proteome. We find that ectopic expression of a single subunit (CCT8) is sufficient to increase TRiC/CCT assembly...
November 28, 2016: Nature Communications
https://www.readbyqxmd.com/read/27890729/maturation-of-human-embryonic-stem-cell-derived-cardiomyocytes-hesc-cms-in-3d-collagen-matrix-effects-of-niche-cell-supplementation-and-mechanical-stimulation
#7
W Zhang, C W Kong, M H Tong, W H Chooi, N Huang, R A Li, B P Chan
: Cardiomyocytes derived from human embryonic stem cells (hESC-CMs) are regarded as a promising source for regenerative medicine, drug testing and disease modeling. Nevertheless, cardiomyocytes are immature in terms of their contractile structure, metabolism and electrophysiological properties. Here, we fabricate cardiac muscle strips by encapsulating hESC-CMs in collagen-based biomaterials. Supplementation of niche cells at 3% to the number of hESC-CMs enhance the maturation of the hESC-CMs in 3D tissue matrix...
November 24, 2016: Acta Biomaterialia
https://www.readbyqxmd.com/read/27888558/spontaneous-single-copy-loss-of-tp53-in-human-embryonic-stem-cells-markedly-increases-cell-proliferation-and-survival
#8
Hadar Amir, Thomas Touboul, Karen Sabatini, Divya Chhabra, Ibon Garitaonandia, Jeanne F Loring, Robert Morey, Louise C Laurent
Genomic aberrations have been identified in many hPSC cultures. Commonly observed duplications in portions of chromosomes 12p and 17q have been associated with increases in genetic instability and resistance to apoptosis, respectively. However, the phenotypic consequences related to sporadic mutations have not been evaluated to date. Here, we report on the effects of a single-copy deletion of the chr17p13.1 region, a sporadic mutation that spontaneously arose independently in several subclones of a hESC culture...
November 26, 2016: Stem Cells
https://www.readbyqxmd.com/read/27879454/molecular-mechanisms-of-androstenediol-in-the-regulation-of-the-proliferative-process-of-human-endometrial-cells
#9
Francisca Plaza-Parrochia, Lorena Oróstica, Paula García, Carolina Vera, Carmen Romero, Luis Valladares, Margarita Vega
Proliferation in endometria of women with polycystic ovarian syndrome (PCOS) is increased, similar to the biosynthesis of androstenediol (estrogenic metabolite). As previously shown, in human endometrial cells, androstenediol increases CYCLIN D1 levels and KI67 and decreases P27 content. The objective of the present investigation was to determine the mechanisms by which androstenediol promotes endometrial cell-cycle progression. Estrogen receptor α (ERα) activation and changes in CYCLIN D1 and P27 levels were evaluated by Western blot in T-HESC and St-T1b endometrial cell lines, using receptor antagonists; activation of PI3K-protein kinase B (AKT) and mitogen-activated protein kinases-extracellular signal-regulated kinases (MAPK-ERK)1/2 pathways was evaluated using PI3K, MAPK/ERK kinase (MEK)1/2, and RNA-polymerase II inhibitors...
November 21, 2016: Reproductive Sciences
https://www.readbyqxmd.com/read/27879221/generation-of-a-tle3-heterozygous-knockout-human-embryonic-stem-cell-line-using-crispr-cas9
#10
Anne M Bara, Angelica Messana, Amanda Herring, Dane Z Hazelbaker, Kevin Eggan, Lindy E Barrett
Here, we generated a monoallelic mutation in the TLE3 (Transducin Like Enhancer of Split 3) gene using CRISPR-Cas9 editing in the human embryonic stem cell (hESC) line WA01. The heterozygous knockout cell line, TLE3-447-D08-A01, displays partial loss of TLE3 protein expression while maintaining pluripotency, differentiation potential and genomic integrity.
September 2016: Stem Cell Research
https://www.readbyqxmd.com/read/27879218/generation-of-a-tle1-homozygous-knockout-human-embryonic-stem-cell-line-using-crispr-cas9
#11
Amanda Herring, Angelica Messana, Anne M Bara, Dane Z Hazelbaker, Kevin Eggan, Lindy E Barrett
Here, we generated a biallelic mutation in the TLE1 (Transducin Like Enhancer of Split 1) gene using CRISPR-Cas9 editing in the human embryonic stem cell (hESC) line WA01. The homozygous knockout cell line, TLE1-464-G04, displays loss of TLE1 protein expression while maintaining pluripotency, differentiation potential and genomic integrity.
September 2016: Stem Cell Research
https://www.readbyqxmd.com/read/27874032/connexin-32-mediated-cell-cell-communication-is-essential-for-hepatic-differentiation-from-human-embryonic-stem-cells
#12
Jinhua Qin, Mingyang Chang, Shuyong Wang, Zhenbo Liu, Wei Zhu, Yi Wang, Fang Yan, Jian Li, Bowen Zhang, Guifang Dou, Jiang Liu, Xuetao Pei, Yunfang Wang
Gap junction-mediated cell-cell interactions are highly conserved and play essential roles in cell survival, proliferation, differentiation and patterning. We report that Connexin 32 (Cx32)-mediated gap junctional intercellular communication (GJIC) is necessary for human embryonic stem cell-derived hepatocytes (hESC-Heps) during step-wise hepatic lineage restriction and maturation. Vitamin K2, previously shown to promote Cx32 expression in mature hepatocytes, up-regulated Cx32 expression and GJIC activation during hepatic differentiation and maturation, resulting in significant increases of hepatic markers expression and hepatocyte functions...
November 22, 2016: Scientific Reports
https://www.readbyqxmd.com/read/27872822/automating-hesc-differentiation-with-3d-printing-and-legacy-liquid-handling-solutions
#13
Eric Zluhan, Kathleen Kelly, Nick LeClair, Danique Wortel, Kesley Moody
Historically, the routine use of laboratory automation solutions has been prohibitively expensive for many laboratories. As legacy hardware has begun to emerge on the secondary market, automation is becoming an increasingly affordable option to augment workflow in virtually any laboratory. To assess the utility of legacy liquid handling in stem cell differentiation, a used liquid handling robot was purchased at auction to automate a stem cell differentiation protocol that gives rise to CD14 + CD45+ mononuclear cells...
2016: MethodsX
https://www.readbyqxmd.com/read/27872482/development-of-a-rapid-screen-for-the-endodermal-differentiation-potential-of-human-pluripotent-stem-cell-lines
#14
Richard Siller, Elena Naumovska, Santosh Mathapati, Max Lycke, Sebastian Greenhough, Gareth J Sullivan
A challenge facing the human pluripotent stem cell (hPSC) field is the variability observed in differentiation potential of hPSCs. Variability can lead to time consuming and costly optimisation to yield the cell type of interest. This is especially relevant for the differentiation of hPSCs towards the endodermal lineages. Endodermal cells have the potential to yield promising new knowledge and therapies for diseases affecting multiple organ systems, including lung, thymus, intestine, pancreas and liver, as well as applications in regenerative medicine and toxicology...
November 22, 2016: Scientific Reports
https://www.readbyqxmd.com/read/27871153/in-vivo-evaluation-of-human-embryonic-stem-cells-isolated-by-57-c11-monoclonal-antibody
#15
Won-Tae Kim, Hyun Min Lee, Min Kyu Kim, Hong Seo Choi, Chun Jeih Ryu
Background: The normal cells derived from human embryonic stem cells (hESCs) are regarded as substitutes for damaged or dysfunctional adult cells. However, tumorigenicity of hESCs remains a major challenge in clinical application of hESC-derived cell transplantation. Previously, we generated monoclonal antibody (MAb) 57-C11 specific to the surface molecule on undifferentiated hESCs. The aim of this study is to prove whether 57-C11-positive hESCs are pluripotent and tumorigenic in immunodeficient mice...
November 30, 2016: International Journal of Stem Cells
https://www.readbyqxmd.com/read/27867005/temporal-expression-of-cd184-cxcr4-and-cd171-l1cam-identifies-distinct-early-developmental-stages-of-human-retinal-ganglion-cells-in-embryonic-stem-cell-derived-retina
#16
J G Aparicio, H Hopp, A Choi, J Mandayam Comar, V C Liao, N Harutyunyan, T C Lee
Human retinal ganglion cells (RGCs) derived from pluripotent stem cells (PSCs) have anticipated value for human disease study, drug screening, and therapeutic applications; however, their full potential remains underdeveloped. To characterize RGCs in human embryonic stem cell (hESC) derived retinal organoids we examined RGC markers and surface antigen expression and made comparisons to human fetal retina. RGCs in both tissues exhibited CD184 and CD171 expression and distinct expression patterns of the RGC markers BRN3 and RBPMS...
November 17, 2016: Experimental Eye Research
https://www.readbyqxmd.com/read/27863459/properties-of-human-embryonic-stem-cells-and-their-differentiated-derivatives-depend-on-non-histone-dna-binding-hmgb1-and-hmgb2-proteins
#17
Alireza J Bagherpoor, Dasa Dolezalova, Tomas Barta, Martin Kucirek, Soodabeh A Sani, Milan Esner, Michaela Kunova Bosakova, Vladimir Vinarsky, Lucie Peskova, Ales Hampl, Michal Stros
HMGB1 and HMGB2 proteins have been implicated in numerous cellular processes, including proliferation, differentiation, apoptosis, and tumor growth. It is unknown whether they are involved in regulating the typical functions of pluripotent human embryonic stem cells (hESCs) and/or those of the differentiated derivatives of hESCs. Using inducible, stably transfected hESCs capable of the shRNA-mediated knockdown of <i>HMGB1</i> and <i>HMGB2</i>, we provide evidence that the downregulation of HMGB1 and/or HMGB2 in undifferentiated hESCs does not affect the stemness of the cells and induces only minor changes to the proliferation rate, cell-cycle profile, and apoptosis...
November 18, 2016: Stem Cells and Development
https://www.readbyqxmd.com/read/27861123/tecrl-a-new-life-threatening-inherited-arrhythmia-gene-associated-with-overlapping-clinical-features-of-both-lqts-and-cpvt
#18
Harsha D Devalla, Roselle Gélinas, Elhadi H Aburawi, Abdelaziz Beqqali, Philippe Goyette, Christian Freund, Marie-A Chaix, Rafik Tadros, Hui Jiang, Antony Le Béchec, Jantine J Monshouwer-Kloots, Tom Zwetsloot, Georgios Kosmidis, Frédéric Latour, Azadeh Alikashani, Maaike Hoekstra, Jurg Schlaepfer, Christine L Mummery, Brian Stevenson, Zoltan Kutalik, Antoine Af de Vries, Léna Rivard, Arthur Am Wilde, Mario Talajic, Arie O Verkerk, Lihadh Al-Gazali, John D Rioux, Zahurul A Bhuiyan, Robert Passier
Genetic causes of many familial arrhythmia syndromes remain elusive. In this study, whole-exome sequencing (WES) was carried out on patients from three different families that presented with life-threatening arrhythmias and high risk of sudden cardiac death (SCD). Two French Canadian probands carried identical homozygous rare variant in TECRL gene (p.Arg196Gln), which encodes the trans-2,3-enoyl-CoA reductase-like protein. Both patients had cardiac arrest, stress-induced atrial and ventricular tachycardia, and QT prolongation on adrenergic stimulation...
December 1, 2016: EMBO Molecular Medicine
https://www.readbyqxmd.com/read/27826002/3d-culture-of-human-pluripotent-stem-cells-in-rgd-alginate-hydrogel-improves-retinal-tissue-development
#19
Nicola C Hunt, Dean Hallam, Ayesha Karimi, Carla B Mellough, Jinju Chen, David H W Steel, Majlinda Lako
: No treatments exist to effectively treat many retinal diseases. Retinal pigmented epithelium (RPE) and neural retina can be generated from human embryonic stem cells/induced pluripotent stem cells (hESCs/hiPSCs). The efficacy of current protocols is, however, limited. It was hypothesised that generation of laminated neural retina and/or RPE from hiPSCs/hESCs could be enhanced by 3D culture in hydrogels. hiPSC- and hESC-derived embryoid bodies (EBs) were encapsulated in 0.5% RGD-alginate; 1% RGD-alginate; hyaluronic acid (HA) or HA/gelatin hydrogels and maintained until day 45...
November 5, 2016: Acta Biomaterialia
https://www.readbyqxmd.com/read/27817157/isoflurane-decreases-proliferation-and-differentiation-but-none-of-the-effects-persist-in-human-embryonic-stem-cell-derived-neural-progenitor-cells
#20
Hye-Min Sohn, Hye Young Kim, Seongjoo Park, Sung-Hee Han, Jin-Hee Kim
PURPOSE: Volatile anesthetics are a potential risk for cognitive impairment in the developing brain. Isoflurane causes cell death, reduces neurogenesis, and changes neuronal differentiation. In this study, the effects of a single isoflurane exposure on the developing human brain were evaluated using human embryonic stem cell (hESC)-derived neural progenitor cells (NPCs). METHODS: Multipotent NPCs were derived from hESCs and randomly exposed to either 5 vol% isoflurane (4 h) or no isoflurane (control group)...
November 5, 2016: Journal of Anesthesia
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