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https://www.readbyqxmd.com/read/29781215/strap-deficiency-impairs-mouse-embryonic-stem-cells-lineage-commitment-through-cyp26a1-mediated-retinoic-acid-homeostasis
#1
Lin Jin, Chenbei Chang, Kevin M Pawlik, Arunima Datta, Larry M Johnson, Trung Vu, Joseph L Napoli, Pran K Datta
Retinoic acid (RA) signaling is essential for the differentiation of embryonic stem cells (ESCs) and vertebrate development. RA biosynthesis and metabolism are controlled by a series of enzymes, but the molecular regulators of these enzymes remain largely obscure. In this study, we investigated the functional role of the WD-domain protein STRAP (serine threonine kinase receptor-associated protein) in the pluripotency and lineage commitment of murine ESCs. We generated Strap knockout (KO) mouse ESCs and subjected them to spontaneous differentiation...
May 21, 2018: Stem Cells
https://www.readbyqxmd.com/read/29765017/assessment-of-established-techniques-to-determine-developmental-and-malignant-potential-of-human-pluripotent-stem-cells
#2
(no author information available yet)
The International Stem Cell Initiative compared several commonly used approaches to assess human pluripotent stem cells (PSC). PluriTest predicts pluripotency through bioinformatic analysis of the transcriptomes of undifferentiated cells, whereas, embryoid body (EB) formation in vitro and teratoma formation in vivo provide direct tests of differentiation. Here we report that EB assays, analyzed after differentiation under neutral conditions and under conditions promoting differentiation to ectoderm, mesoderm, or endoderm lineages, are sufficient to assess the differentiation potential of PSCs...
May 15, 2018: Nature Communications
https://www.readbyqxmd.com/read/29761384/a-simple-multistep-protocol-for-differentiating-human-induced-pluripotent-stem-cells-into-functional-macrophages
#3
Chandrayana Mukherjee, Christine Hale, Subhankar Mukhopadhyay
Macrophages differentiated from human induced pluripotent stem cells (hiPSCs) provide an alternative new tool overcoming some of the limitations of existing models for human macrophages, such as human macrophage-like cell lines and primary monocyte-derived macrophages. A combination of different cytokines and growth factors can differentiate hiPSCs toward myeloid lineage. Here we describe a simple multistep protocol for differentiating hiPSCs into functional macrophages. This includes derivation of three germ-line containing embryoid bodies (EBs) from iPSCs, generation of myeloid precursors from EBs, and finally maturation of myeloid precursors into functional macrophages...
2018: Methods in Molecular Biology
https://www.readbyqxmd.com/read/29734353/knockdown-of-cdk2ap1-in-human-embryonic-stem-cells-reduces-the-threshold-of-differentiation
#4
Khaled N Alsayegh, Steven D Sheridan, Shilpa Iyer, Raj Raghavendra Rao
Recent studies have suggested a role for the Cyclin Dependent Kinase-2 Associated Protein 1 (CDK2AP1) in stem cell differentiation and self-renewal. In studies with mouse embryonic stem cells (mESCs) derived from generated mice embryos with targeted deletion of the Cdk2ap1 gene, CDK2AP1 was shown to be required for epigenetic silencing of Oct4 during differentiation, with deletion resulting in persistent self-renewal and reduced differentiation potential. Differentiation capacity was restored in these cells following the introduction of a non-phosphorylatible form of the retinoblastoma protein (pRb) or exogenous Cdk2ap1...
2018: PloS One
https://www.readbyqxmd.com/read/29670257/directed-reprogramming-of-comprehensively-characterized-dental-pulp-stem-cells-extracted-from-natal-tooth
#5
Rishikaysh V Pisal, Jakub Suchanek, Richard Siller, Tomas Soukup, Hana Hrebikova, Ales Bezrouk, David Kunke, Stanislav Micuda, Stanislav Filip, Gareth Sullivan, Jaroslav Mokry
The aim of this study was to extensively characterise natal dental pulp stem cells (nDPSC) and assess their efficiency to generate human induced pluripotent stem cells (hiPSC). A number of distinguishing features prompted us to choose nDPSC over normal adult DPSC, in that they differed in cell surface marker expression and initial doubling time. In addition, nDPSC expressed 17 out of 52 pluripotency genes we analysed, and the level of expression was comparable to human embryonic stem cells (hESC). Ours is the first group to report comprehensive characterization of nDPSC followed by directed reprogramming to a pluripotent stem cell state...
April 18, 2018: Scientific Reports
https://www.readbyqxmd.com/read/29666052/functional-comparison-of-distinct-brachyury-states-in-a-renal-differentiation-assay
#6
Jing Zhou, Antonius Plagge, Patricia Murray
Mesodermal populations can be generated in vitro from mouse embryonic stem cells (mESCs) using three-dimensional (3-D) aggregates called embryoid bodies or two-dimensional (2-D) monolayer culture systems. Here, we investigated whether Brachyury -expressing mesodermal cells generated using 3-D or 2-D culture systems are equivalent, or instead, have different properties. Using a Brachyury -GFP/E2-Crimson reporter mESC line, we isolated Brachyury- GFP+ mesoderm cells using flow-activated cell sorting and compared their gene expression profiles and ex vivo differentiation patterns...
April 17, 2018: Biology Open
https://www.readbyqxmd.com/read/29664925/actin-and-myosin-ii-modulate-differentiation-of-pluripotent-stem-cells
#7
Liana C Boraas, Emma T Pineda, Tabassum Ahsan
Use of stem cell-based therapies in tissue engineering and regenerative medicine is hindered by efficient means of directed differentiation. For pluripotent stem cells, an initial critical differentiation event is specification to one of three germ lineages: endoderm, mesoderm, and ectoderm. Differentiation is known to be regulated by numerous extracellular and intracellular factors, but the role of the cytoskeleton during specification, or early differentiation, is still unknown. In these studies, we used agonists and antagonists to modulate actin polymerization and the actin-myosin molecular motor during spontaneous differentiation of embryonic stem cells in embryoid bodies...
2018: PloS One
https://www.readbyqxmd.com/read/29658075/induced-pluripotent-stem-cell-derived-hematopoietic-embryoid-bodies-secrete-sphingosine-1-phosphate-and-revert-endothelial-injury
#8
S Kasuda, R Kudo, K Yuui, Y Sakurai, K Hatake
The possibility of sphingosine-1-phosphate production by induced pluripotent stem cells is examined to assess their potential in treatment of sepsis. The hematopoietic embryoid bodies were derived from the culture of 6-day-old differentiated induced pluripotent stem cells. These embryoid bodies secreted sphingosine-1-phosphate, an important bioactive lipid that regulates integrity of the pulmonary endothelial barrier, prevents elevation of its permeability, and impedes the formation of stress fibers in human endotheliocytes derived from umbilical vein...
April 16, 2018: Bulletin of Experimental Biology and Medicine
https://www.readbyqxmd.com/read/29622030/mapping-human-pluripotent-stem-cell-differentiation-pathways-using-high-throughput-single-cell-rna-sequencing
#9
Xiaoping Han, Haide Chen, Daosheng Huang, Huidong Chen, Lijiang Fei, Chen Cheng, He Huang, Guo-Cheng Yuan, Guoji Guo
BACKGROUND: Human pluripotent stem cells (hPSCs) provide powerful models for studying cellular differentiations and unlimited sources of cells for regenerative medicine. However, a comprehensive single-cell level differentiation roadmap for hPSCs has not been achieved. RESULTS: We use high throughput single-cell RNA-sequencing (scRNA-seq), based on optimized microfluidic circuits, to profile early differentiation lineages in the human embryoid body system. We present a cellular-state landscape for hPSC early differentiation that covers multiple cellular lineages, including neural, muscle, endothelial, stromal, liver, and epithelial cells...
April 5, 2018: Genome Biology
https://www.readbyqxmd.com/read/29602048/lymphoblast-derived-integration-free-ipsc-line-ad-trem2-1-from-a-67year-old-alzheimer-s-disease-patient-expressing-the-trem2-p-r47h-variant
#10
Soraia Martins, Hatice Yigit, Martina Bohndorf, Nina Graffmann, Aurelian Robert Fiszl, Wasco Wruck, Kristel Sleegers, Christine Van Broeckhoven, James Adjaye
Human lymphoblast cells from a male diagnosed with Alzheimer's disease (AD) expressing the TREM2 p.R47H variant were used to generate integration-free induced pluripotent stem cells (iPSCs) by over-expressing episomal-based plasmids harbouring OCT4, SOX2, NANOG, LIN28, c-MYC and L-MYC. AD-TREM2-1 was defined as pluripotent based on (i) expression of pluripotency-associated markers (ii) embryoid body-based differentiation into cell types representative of the three germ layers and (iii) the similarity between the transcriptome of the iPSC line and the human embryonic stem cell line H1 with a Pearson correlation of 0...
March 20, 2018: Stem Cell Research
https://www.readbyqxmd.com/read/29576547/expression-specificity-and-compensation-effect-of-ash2l-1-ash2l-2-in-mouse-embryonic-stem-cells
#11
Jing Xie, Chen Fan, Jing-Long Zhang, Shi-Qiang Zhang
H3K4me3 is an important epigenetic modification that plays a critical role in maintaining self-renewal of mouse embryonic stem cells (mESCs). H3K4me3 is catalyzed mainly by the mixed lineage leukemia (MLL) methyl-transferase complex. ASH2L, a core subunit of the MLL complex, participates in regulating the open state of chromatin in mESCs. There are two isoforms of the ASH2L protein: ASH2L-1 (80 kDa), which only exists in mouse embryonic fibroblasts and ASH2L-2 (65 kDa), which is the predominant isoform in mESCs...
March 20, 2018: Yi Chuan, Hereditas
https://www.readbyqxmd.com/read/29555651/an-inactivating-mutation-in-the-histone-deacetylase-sirt6-causes-human-perinatal-lethality
#12
Christina M Ferrer, Marielle Alders, Alex V Postma, Seonmi Park, Mark A Klein, Murat Cetinbas, Eva Pajkrt, Astrid Glas, Silvana van Koningsbruggen, Vincent M Christoffels, Marcel M A M Mannens, Lia Knegt, Jean-Pierre Etchegaray, Ruslan I Sadreyev, John M Denu, Gustavo Mostoslavsky, Merel C van Maarle, Raul Mostoslavsky
It has been well established that histone and DNA modifications are critical to maintaining the equilibrium between pluripotency and differentiation during early embryogenesis. Mutations in key regulators of DNA methylation have shown that the balance between gene regulation and function is critical during neural development in early years of life. However, there have been no identified cases linking epigenetic regulators to aberrant human development and fetal demise. Here, we demonstrate that a homozygous inactivating mutation in the histone deacetylase SIRT6 results in severe congenital anomalies and perinatal lethality in four affected fetuses...
March 1, 2018: Genes & Development
https://www.readbyqxmd.com/read/29554331/in-vitro-platform-of-allogeneic-stem-cell-derived-cardiomyocyte-transplantation-for-cardiac-conduction-defects
#13
Akira Yoshida, Jong-Kook Lee, Satoki Tomoyama, Keiko Miwa, Keiichi Shirakawa, Sanae Hamanaka, Tomoyuki Yamaguchi, Hiromitsu Nakauchi, Shigeru Miyagawa, Yoshiki Sawa, Issei Komuro, Yasushi Sakata
Aims: The aim of the present study is to develop in vitro experimental analytical method for the electrophysiological properties of allogeneic induced pluripotent stem cell-derived cardiomyocytes (CMs) in cardiac conduction defect model. Methods and results: Cardiomyocytes were derived from rat induced pluripotent stem cells CMs (riPSC-CMs) using an embryoid body-based differentiation method with the serial application of growth factors including activin-A, bone morphogenetic protein 4 (BMP-4), and inhibitor of wnt production 2 (IWP-2)...
March 15, 2018: Europace: European Pacing, Arrhythmias, and Cardiac Electrophysiology
https://www.readbyqxmd.com/read/29546877/hsp60-is-required-for-stemness-and-proper-differentiation-of-mouse-embryonic-stem-cells
#14
Nan-Hee Seo, Eun-Hye Lee, Jin-Hee Seo, Hwa-Ryung Song, Myung-Kwan Han
Embryonic stem cells (ESCs) are metabolically distinct from their differentiated counterparts. ESC mitochondria are less complex and fewer in number than their differentiated progeny. However, few studies have examined the proteins responsible for differences in mitochondrial structure and function between ESCs and somatic cells. Therefore, in this study, we aimed to investigate the differences between mitochondrial proteins in these two cell types. We demonstrate that HSP60 is more abundant in mouse ESC mitochondria than in mouse embryonic fibroblasts...
March 16, 2018: Experimental & Molecular Medicine
https://www.readbyqxmd.com/read/29544541/highly-efficient-methods-to-obtain-homogeneous-dorsal-neural-progenitor-cells-from-human-and-mouse-embryonic-stem-cells-and-induced-pluripotent-stem-cells
#15
Meixiang Zhang, Justine Ngo, Filomena Pirozzi, Ying-Pu Sun, Anthony Wynshaw-Boris
BACKGROUND: Embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) have been widely used to generate cellular models harboring specific disease-related genotypes. Of particular importance are ESC and iPSC applications capable of producing dorsal telencephalic neural progenitor cells (NPCs) that are representative of the cerebral cortex and overcome the challenges of maintaining a homogeneous population of cortical progenitors over several passages in vitro. While previous studies were able to derive NPCs from pluripotent cell types, the fraction of dorsal NPCs in this population is small and decreases over several passages...
March 15, 2018: Stem Cell Research & Therapy
https://www.readbyqxmd.com/read/29543795/generation-of-spatial-patterned-early-developing-cardiac-organoids-using-human-pluripotent-stem-cells
#16
Plansky Hoang, Jason Wang, Bruce R Conklin, Kevin E Healy, Zhen Ma
The creation of human induced pluripotent stem cells (hiPSCs) has provided an unprecedented opportunity to study tissue morphogenesis and organ development through 'organogenesis-in-a-dish'. Current approaches to cardiac organoid engineering rely on either direct cardiac differentiation from embryoid bodies (EBs) or generation of aligned cardiac tissues from predifferentiated cardiomyocytes from monolayer hiPSCs. To experimentally model early cardiac organogenesis in vitro, our protocol combines biomaterials-based cell patterning with stem cell organoid engineering...
April 2018: Nature Protocols
https://www.readbyqxmd.com/read/29540665/myocardial-bmp2-gain-causes-ectopic-emt-and-promotes-cardiomyocyte-proliferation-and-immaturity
#17
Belén Prados, Paula Gómez-Apiñániz, Tania Papoutsi, Guillermo Luxán, Stephane Zaffran, José María Pérez-Pomares, José Luis de la Pompa
During mammalian heart development, restricted myocardial Bmp2 expression is a key patterning signal for atrioventricular canal specification and the epithelial-mesenchyme transition that gives rise to the valves. Using a mouse transgenic line conditionally expressing Bmp2, we show that widespread Bmp2 expression in the myocardium leads to valve and chamber dysmorphogenesis and embryonic death by E15.5. Transgenic embryos show thickened valves, ventricular septal defect, enlarged trabeculae and dilated ventricles, with an endocardium able to undergo EMT both in vivo and in vitro...
March 14, 2018: Cell Death & Disease
https://www.readbyqxmd.com/read/29531836/mitochondrial-fission-protein-drp1-inhibition-promotes-cardiac-mesodermal-differentiation-of-human-pluripotent-stem-cells
#18
Ashfaqul Hoque, Priyadharshini Sivakumaran, Simon T Bond, Naomi X Y Ling, Anne M Kong, John W Scott, Nadeeka Bandara, Damián Hernández, Guei-Sheung Liu, Raymond C B Wong, Michael T Ryan, Derek J Hausenloy, Bruce E Kemp, Jonathan S Oakhill, Brian G Drew, Alice Pébay, Shiang Y Lim
Human induced pluripotent stem cells (iPSCs) are a valuable tool for studying the cardiac developmental process in vitro, and cardiomyocytes derived from iPSCs are a putative cell source for personalized medicine. Changes in mitochondrial morphology have been shown to occur during cellular reprogramming and pluripotent stem cell differentiation. However, the relationships between mitochondrial dynamics and cardiac mesoderm commitment of iPSCs remain unclear. Here we demonstrate that changes in mitochondrial morphology from a small granular fragmented phenotype in pluripotent stem cells to a filamentous reticular elongated network in differentiated cardiomyocytes are required for cardiac mesodermal differentiation...
December 2018: Cell Death Discovery
https://www.readbyqxmd.com/read/29513687/generation-of-functional-cardiomyocytes-from-rat-embryonic-and-induced-pluripotent-stem-cells-using-feeder-free-expansion-and-differentiation-in-suspension-culture
#19
Julia Dahlmann, George Awad, Carsten Dolny, Sönke Weinert, Karin Richter, Klaus-Dieter Fischer, Thomas Munsch, Volkmar Leßmann, Marianne Volleth, Martin Zenker, Yaoyao Chen, Claudia Merkl, Angelika Schnieke, Hassina Baraki, Ingo Kutschka, George Kensah
The possibility to generate cardiomyocytes from pluripotent stem cells in vitro has enormous significance for basic research, disease modeling, drug development and heart repair. The concept of heart muscle reconstruction has been studied and optimized in the rat model using rat primary cardiovascular cells or xenogeneic pluripotent stem cell derived-cardiomyocytes for years. However, the lack of rat pluripotent stem cells (rPSCs) and their cardiovascular derivatives prevented the establishment of an authentic clinically relevant syngeneic or allogeneic rat heart regeneration model...
2018: PloS One
https://www.readbyqxmd.com/read/29512128/use-of-human-embryoid-bodies-for-teratology
#20
Anthony Flamier, Supriya Singh, Theodore P Rasmussen
Human birth defects are relatively common and can be caused by exposure to environmental teratogens or to pharmaceuticals with teratogenic activities. Human embryonic stem cells (hESCs), by virtue of their pluripotent nature, provide an excellent cellular platform for teratogen detection and risk assessment. This unit describes detailed protocols for the preparation and validation of highly pluripotent hESCs, the production of large quantities of aggregated multicellular spheroids composed of hESCs, and these spheroids' differentiation into embryoid bodies (EBs)...
February 21, 2018: Current Protocols in Toxicology
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