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Stem Cell Reports

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https://www.readbyqxmd.com/read/29779896/all-optical-electrophysiology-for-high-throughput-functional-characterization-of-a-human-ipsc-derived-motor-neuron-model-of-als
#1
Evangelos Kiskinis, Joel M Kralj, Peng Zou, Eli N Weinstein, Hongkang Zhang, Konstantinos Tsioras, Ole Wiskow, J Alberto Ortega, Kevin Eggan, Adam E Cohen
Human induced pluripotent stem cell (iPSC)-derived neurons are an attractive substrate for modeling disease, yet the heterogeneity of these cultures presents a challenge for functional characterization by manual patch-clamp electrophysiology. Here, we describe an optimized all-optical electrophysiology, "Optopatch," pipeline for high-throughput functional characterization of human iPSC-derived neuronal cultures. We demonstrate the method in a human iPSC-derived motor neuron (iPSC-MN) model of amyotrophic lateral sclerosis (ALS)...
May 11, 2018: Stem Cell Reports
https://www.readbyqxmd.com/read/29779899/signaling-dependent-control-of-apical-membrane-size-and-self-renewal-in-rosette-stage-human-neuroepithelial-stem-cells
#2
Jan-Philip Medelnik, Kathleen Roensch, Satoshi Okawa, Antonio Del Sol, Osvaldo Chara, Levan Mchedlishvili, Elly M Tanaka
In the developing nervous system, neural stem cells are polarized and maintain an apical domain facing a central lumen. The presence of apical membrane is thought to have a profound influence on maintaining the stem cell state. With the onset of neurogenesis, cells lose their polarization, and the concomitant loss of the apical domain coincides with a loss of the stem cell identity. Little is known about the molecular signals controlling apical membrane size. Here, we use two neuroepithelial cell systems, one derived from regenerating axolotl spinal cord and the other from human embryonic stem cells, to identify a molecular signaling pathway initiated by lysophosphatidic acid that controls apical membrane size and consequently controls and maintains epithelial organization and lumen size in neuroepithelial rosettes...
May 10, 2018: Stem Cell Reports
https://www.readbyqxmd.com/read/29779898/oncostatin-m-and-kit-ligand-control-hematopoietic-stem-cell-fate-during-zebrafish-embryogenesis
#3
Christopher B Mahony, Corentin Pasche, Julien Y Bertrand
Understanding the molecular pathways controlling hematopoietic stem cell specification and expansion is a necessary milestone to perform regenerative medicine. Here, we used the zebrafish model to study the role of the ckit signaling pathway in this process. We show the importance of kitb/kitlgb signaling in the specification and expansion of hematopoietic stem cells (HSCs), in the hemogenic endothelium and caudal hematopoietic tissue (CHT), respectively. Moreover, we identified the zebrafish ortholog of Oncostatin M (osm) in the zebrafish genome...
May 10, 2018: Stem Cell Reports
https://www.readbyqxmd.com/read/29779897/inductive-and-selective-effects-of-gsk3-and-mek-inhibition-on-nanog-heterogeneity-in-embryonic-stem-cells
#4
Simon Hastreiter, Stavroula Skylaki, Dirk Loeffler, Andreas Reimann, Oliver Hilsenbeck, Philipp S Hoppe, Daniel L Coutu, Konstantinos D Kokkaliaris, Michael Schwarzfischer, Konstantinos Anastassiadis, Fabian J Theis, Timm Schroeder
Embryonic stem cells (ESCs) display heterogeneous expression of pluripotency factors such as Nanog when cultured with serum and leukemia inhibitory factor (LIF). In contrast, dual inhibition of the signaling kinases GSK3 and MEK (2i) converts ESC cultures into a state with more uniform and high Nanog expression. However, it is so far unclear whether 2i acts through an inductive or selective mechanism. Here, we use continuous time-lapse imaging to quantify the dynamics of death, proliferation, and Nanog expression in mouse ESCs after 2i addition...
May 10, 2018: Stem Cell Reports
https://www.readbyqxmd.com/read/29779895/identification-and-single-cell-functional-characterization-of-an-endodermally-biased-pluripotent-substate-in-human-embryonic-stem-cells
#5
Thomas F Allison, Andrew J H Smith, Konstantinos Anastassiadis, Jackie Sloane-Stanley, Veronica Biga, Dylan Stavish, James Hackland, Shan Sabri, Justin Langerman, Mark Jones, Kathrin Plath, Daniel Coca, Ivana Barbaric, Paul Gokhale, Peter W Andrews
Human embryonic stem cells (hESCs) display substantial heterogeneity in gene expression, implying the existence of discrete substates within the stem cell compartment. To determine whether these substates impact fate decisions of hESCs we used a GFP reporter line to investigate the properties of fractions of putative undifferentiated cells defined by their differential expression of the endoderm transcription factor, GATA6, together with the hESC surface marker, SSEA3. By single-cell cloning, we confirmed that substates characterized by expression of GATA6 and SSEA3 include pluripotent stem cells capable of long-term self-renewal...
May 9, 2018: Stem Cell Reports
https://www.readbyqxmd.com/read/29779894/epigenetic-regulation-by-baf-complexes-limits-neural-stem-cell-proliferation-by-suppressing-wnt-signaling-in-late-embryonic-development
#6
Huong Nguyen, Cemil Kerimoglu, Mehdi Pirouz, Linh Pham, Kamila A Kiszka, Godwin Sokpor, M Sadman Sakib, Joachim Rosenbusch, Ulrike Teichmann, Rho H Seong, Anastassia Stoykova, Andre Fischer, Jochen F Staiger, Tran Tuoc
During early cortical development, neural stem cells (NSCs) divide symmetrically to expand the progenitor pool, whereas, in later stages, NSCs divide asymmetrically to self-renew and produce other cell types. The timely switch from such proliferative to differentiative division critically determines progenitor and neuron numbers. However, the mechanisms that limit proliferative division in late cortical development are not fully understood. Here, we show that the BAF (mSWI/SNF) complexes restrict proliferative competence and promote neuronal differentiation in late corticogenesis...
May 8, 2018: Stem Cell Reports
https://www.readbyqxmd.com/read/29754959/human-induced-pluripotent-stem-cell-derived-cardiac-cell-sheets-expressing-genetically-encoded-voltage-indicator-for-pharmacological-and-arrhythmia-studies
#7
Naim Shaheen, Assad Shiti, Irit Huber, Rami Shinnawi, Gil Arbel, Amira Gepstein, Noga Setter, Idit Goldfracht, Amit Gruber, Snizhanna V Chorna, Lior Gepstein
Fulfilling the potential of human induced pluripotent stem cell (hiPSC)-derived cardiomyocytes for studying conduction and arrhythmogenesis requires development of multicellular models and methods for long-term repeated tissue phenotyping. We generated confluent hiPSC-derived cardiac cell sheets (hiPSC-CCSs), expressing the genetically encoded voltage indicator ArcLight. ArcLight-based optical mapping allowed generation of activation and action-potential duration (APD) maps, which were validated by mapping the same hiPSC-CCSs with the voltage-sensitive dye, Di-4-ANBDQBS...
May 7, 2018: Stem Cell Reports
https://www.readbyqxmd.com/read/29754961/hematopoietic-stem-cells-but-not-multipotent-progenitors-drive-erythropoiesis-during-chronic-erythroid-stress-in-epo-transgenic-mice
#8
Rashim Pal Singh, Tatyana Grinenko, Beáta Ramasz, Kristin Franke, Mathias Lesche, Andreas Dahl, Max Gassmann, Triantafyllos Chavakis, Ian Henry, Ben Wielockx
The hematopoietic stem cell (HSC) compartment consists of a small pool of cells capable of replenishing all blood cells. Although it is established that the hematopoietic system is assembled as a hierarchical organization under steady-state conditions, emerging evidence suggests that distinct differentiation pathways may exist in response to acute stress. However, it remains unclear how different hematopoietic stem and progenitor cell subpopulations behave under sustained chronic stress. Here, by using adult transgenic mice overexpressing erythropoietin (EPO; Tg6) and a combination of in vivo, in vitro, and deep-sequencing approaches, we found that HSCs respond differentially to chronic erythroid stress compared with their closely related multipotent progenitors (MPPs)...
May 4, 2018: Stem Cell Reports
https://www.readbyqxmd.com/read/29754960/high-level-precise-knockin-of-ipscs-by-simultaneous-reprogramming-and-genome-editing-of-human-peripheral-blood-mononuclear-cells
#9
Wei Wen, Xinxin Cheng, Yawen Fu, Feiying Meng, Jian-Ping Zhang, Lu Zhang, Xiao-Lan Li, Zhixue Yang, Jing Xu, Feng Zhang, Gary D Botimer, Weiping Yuan, Changkai Sun, Tao Cheng, Xiao-Bing Zhang
We have developed an improved episomal vector system for efficient generation of integration-free induced pluripotent stem cells (iPSCs) from peripheral blood mononuclear cells. More recently, we reported that the use of an optimized CRISPR-Cas9 system together with a double-cut donor increases homology-directed repair-mediated precise gene knockin efficiency by 5- to 10-fold. Here, we report the integration of blood cell reprogramming and genome editing in a single step. We found that expression of Cas9 and KLF4 using a single vector significantly increases genome editing efficiency, and addition of SV40LT further enhances knockin efficiency...
May 4, 2018: Stem Cell Reports
https://www.readbyqxmd.com/read/29754958/automated-deep-learning-based-system-to-identify-endothelial-cells-derived-from-induced-pluripotent-stem-cells
#10
Dai Kusumoto, Mark Lachmann, Takeshi Kunihiro, Shinsuke Yuasa, Yoshikazu Kishino, Mai Kimura, Toshiomi Katsuki, Shogo Itoh, Tomohisa Seki, Keiichi Fukuda
Deep learning technology is rapidly advancing and is now used to solve complex problems. Here, we used deep learning in convolutional neural networks to establish an automated method to identify endothelial cells derived from induced pluripotent stem cells (iPSCs), without the need for immunostaining or lineage tracing. Networks were trained to predict whether phase-contrast images contain endothelial cells based on morphology only. Predictions were validated by comparison to immunofluorescence staining for CD31, a marker of endothelial cells...
May 3, 2018: Stem Cell Reports
https://www.readbyqxmd.com/read/29731430/pluripotent-stem-cell-model-of-nakajo-nishimura-syndrome-untangles-proinflammatory-pathways-mediated-by-oxidative-stress
#11
Fumiko Honda-Ozaki, Madoka Terashima, Akira Niwa, Norikazu Saiki, Yuri Kawasaki, Haruna Ito, Akitsu Hotta, Ayako Nagahashi, Koichi Igura, Isao Asaka, Hongmei Lisa Li, Masakatsu Yanagimachi, Fukumi Furukawa, Nobuo Kanazawa, Tatsutoshi Nakahata, Megumu K Saito
Nakajo-Nishimura syndrome (NNS) is an immunoproteasome-associated autoinflammatory disorder caused by a mutation of the PSMB8 gene. Although dysfunction of the immunoproteasome causes various cellular stresses attributed to the overproduction of inflammatory cytokines and chemokines in NNS, the underlying mechanisms of the autoinflammation are still largely unknown. To investigate and understand the mechanisms and signal pathways in NNS, we established a panel of isogenic pluripotent stem cell (PSC) lines with PSMB8 mutation...
May 1, 2018: Stem Cell Reports
https://www.readbyqxmd.com/read/29731431/large-scale-expansion-of-human-ipsc-derived-skeletal-muscle-cells-for-disease-modeling-and-cell-based-therapeutic-strategies
#12
Erik van der Wal, Pablo Herrero-Hernandez, Raymond Wan, Mike Broeders, Stijn L M In 't Groen, Tom J M van Gestel, Wilfred F J van IJcken, Tom H Cheung, Ans T van der Ploeg, Gerben J Schaaf, W W M Pim Pijnappel
Although skeletal muscle cells can be generated from human induced pluripotent stem cells (iPSCs), transgene-free protocols include only limited options for their purification and expansion. In this study, we found that fluorescence-activated cell sorting-purified myogenic progenitors generated from healthy controls and Pompe disease iPSCs can be robustly expanded as much as 5 × 1011 -fold. At all steps during expansion, cells could be cryopreserved or differentiated into myotubes with a high fusion index...
April 28, 2018: Stem Cell Reports
https://www.readbyqxmd.com/read/29731429/electrophysiologic-characterization-of-calcium-handling-in-human-induced-pluripotent-stem-cell-derived-atrial-cardiomyocytes
#13
Mariana Argenziano, Erin Lambers, Liang Hong, Arvind Sridhar, Meihong Zhang, Brandon Chalazan, Ambili Menon, Eleonora Savio-Galimberti, Joseph C Wu, Jalees Rehman, Dawood Darbar
Human induced pluripotent stem cell (hiPSC)-derived atrial cardiomyocytes (CMs) hold great promise for elucidating underlying cellular mechanisms that cause atrial fibrillation (AF). In order to use atrial-like hiPSC-CMs for arrhythmia modeling, it is essential to better understand the molecular and electrophysiological phenotype of these cells. We performed comprehensive molecular, transcriptomic, and electrophysiologic analyses of retinoic acid (RA)-guided hiPSC atrial-like CMs and demonstrate that RA results in differential expression of genes involved in calcium ion homeostasis that directly interact with an RA receptor, chicken ovalbumin upstream promoter-transcription factor 2 (COUP-TFII)...
April 28, 2018: Stem Cell Reports
https://www.readbyqxmd.com/read/29706500/thyroid-hormone-transporters-mct8-and-oatp1c1-control-skeletal-muscle-regeneration
#14
Steffen Mayerl, Manuel Schmidt, Denica Doycheva, Veerle M Darras, Sören S Hüttner, Anita Boelen, Theo J Visser, Christoph Kaether, Heike Heuer, Julia von Maltzahn
Thyroid hormone (TH) transporters are required for the transmembrane passage of TH in target cells. In humans, inactivating mutations in the TH transporter MCT8 cause the Allan-Herndon-Dudley syndrome, characterized by severe neuromuscular symptoms and an abnormal TH serum profile, which is fully replicated in Mct8 knockout mice and Mct8/Oatp1c1 double-knockout (M/O DKO) mice. Analysis of tissue TH content and expression of TH-regulated genes indicate a thyrotoxic state in Mct8-deficient skeletal muscles. Both TH transporters are upregulated in activated satellite cells (SCs)...
April 25, 2018: Stem Cell Reports
https://www.readbyqxmd.com/read/29706501/inducible-expression-of-gdnf-in-transplanted-ipsc-derived-neural-progenitor-cells
#15
Aslam Abbasi Akhtar, Genevieve Gowing, Naomi Kobritz, Steve E Savinoff, Leslie Garcia, David Saxon, Noell Cho, Gibum Kim, Colton M Tom, Hannah Park, George Lawless, Brandon C Shelley, Virginia B Mattis, Joshua J Breunig, Clive N Svendsen
Trophic factor delivery to the brain using stem cell-derived neural progenitors is a powerful way to bypass the blood-brain barrier. Protection of diseased neurons using this technology is a promising therapy for neurodegenerative diseases. Glial cell line-derived neurotrophic factor (GDNF) has provided benefits to Parkinsonian patients and is being used in a clinical trial for amyotrophic lateral sclerosis. However, chronic trophic factor delivery prohibits dose adjustment or cessation if side effects develop...
April 23, 2018: Stem Cell Reports
https://www.readbyqxmd.com/read/29706502/unraveling-the-inconsistencies-of-cardiac-differentiation-efficiency-induced-by-the-gsk3%C3%AE-inhibitor-chir99021-in-human-pluripotent-stem-cells
#16
Filip Laco, Tsung Liang Woo, Qixing Zhong, Radoslaw Szmyd, Sherwin Ting, Fahima Jaleel Khan, Christina L L Chai, Shaul Reuveny, Allen Chen, Steve Oh
Cardiac differentiation efficiency is hampered by inconsistencies and low reproducibility. We analyzed the differentiation process of multiple human pluripotent stem cell (hPSC) lines in response to dynamic GSK3β inhibition under varying cell culture conditions. hPSCs showed strong differences in cell-cycle profiles with varying culture confluency. hPSCs with a higher percentage of cells in the G1 phase of the cell cycle exhibited cell death and required lower doses of GSK3β inhibitors to induce cardiac differentiation...
April 20, 2018: Stem Cell Reports
https://www.readbyqxmd.com/read/29706499/ink4a-arf-expression-impairs-neurogenesis-in-the-brain-of-irradiated-mice
#17
Oanh Le, Lina Palacio, Gilbert Bernier, Ines Batinic-Haberle, Gilles Hickson, Christian Beauséjour
Brain neurogenesis is severely impaired following exposure to ionizing radiation (IR). We and others have shown that the expression of the tumor suppressor gene p16INK4a is increased in tissues exposed to IR and thus hypothesized that its expression could limit neurogenesis in the irradiated brain. Here, we found that exposure to IR leads to persistent DNA damage and the expression of p16INK4a in the hippocampus and subventricular zone regions. This was accompanied by a decline in neurogenesis, as determined by doublecortin expression and bromodeoxyuridine incorporation, an effect partially restored in Ink4a/arf-null mice...
April 20, 2018: Stem Cell Reports
https://www.readbyqxmd.com/read/29706498/the-stat3-target-mettl8-regulates-mouse-esc-differentiation-via-inhibiting-the-jnk-pathway
#18
Hao Gu, Dang Vinh Do, Xinyu Liu, Luang Xu, Yixun Su, Jie Min Nah, Yuqian Wong, Ying Li, Na Sheng, Gebreselassie Addisu Tilaye, Henry Yang, Huili Guo, Jun Yan, Xin-Yuan Fu
The capacity of embryonic stem cells (ESCs) to differentiate into all lineages of mature organism is precisely regulated by cellular signaling factors. STAT3 is a crucial transcription factor that plays a central role in maintaining ESC identity. However, the underlying mechanism by which STAT3 directs differentiation is still not completely understood. Here, we show that STAT3 positively regulates gene expression of methyltransferase-like protein 8 (Mettl8) in mouse ESCs. We found that METTL8 is dispensable for pluripotency but affects ESC differentiation...
April 20, 2018: Stem Cell Reports
https://www.readbyqxmd.com/read/29681545/nf-%C3%AE%C2%BAb-activity-initiates-human-esc-derived-neural-progenitor-cell-differentiation-by-inducing-a-metabolic-maturation-program
#19
Lorna M FitzPatrick, Kate E Hawkins, Juliette M K M Delhove, Emilio Fernandez, Chiara Soldati, Louise F Bullen, Axel Nohturfft, Simon N Waddington, Diego L Medina, Juan P Bolaños, Tristan R McKay
Human neural development begins at embryonic day 19 and marks the beginning of organogenesis. Neural stem cells in the neural tube undergo profound functional, morphological, and metabolic changes during neural specification, coordinated by a combination of exogenous and endogenous cues. The temporal cell signaling activities that mediate this process, during development and in the postnatal brain, are incompletely understood. We have applied gene expression studies and transcription factor-activated reporter lentiviruses during in vitro neural specification of human pluripotent stem cells...
April 17, 2018: Stem Cell Reports
https://www.readbyqxmd.com/read/29681544/dissecting-the-contributions-of-cooperating-gene-mutations-to-cancer-phenotypes-and-drug-responses-with-patient-derived-ipscs
#20
Chan-Jung Chang, Andriana G Kotini, Malgorzata Olszewska, Maria Georgomanoli, Julie Teruya-Feldstein, Henrik Sperber, Roberto Sanchez, Robert DeVita, Timothy J Martins, Omar Abdel-Wahab, Robert K Bradley, Eirini P Papapetrou
Connecting specific cancer genotypes with phenotypes and drug responses constitutes the central premise of precision oncology but is hindered by the genetic complexity and heterogeneity of primary cancer cells. Here, we use patient-derived induced pluripotent stem cells (iPSCs) and CRISPR/Cas9 genome editing to dissect the individual contributions of two recurrent genetic lesions, the splicing factor SRSF2 P95L mutation and the chromosome 7q deletion, to the development of myeloid malignancy. Using a comprehensive panel of isogenic iPSCs-with none, one, or both genetic lesions-we characterize their relative phenotypic contributions and identify drug sensitivities specific to each one through a candidate drug approach and an unbiased large-scale small-molecule screen...
April 17, 2018: Stem Cell Reports
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