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Human embryonic stem cells derived cardiomyocytes

Natalie Weber, Kristin Schwanke, Stephan Greten, Meike Wendland, Bogdan Iorga, Martin Fischer, Cornelia Geers-Knörr, Jan Hegermann, Christoph Wrede, Jan Fiedler, Henning Kempf, Annika Franke, Birgit Piep, Angelika Pfanne, Thomas Thum, Ulrich Martin, Bernhard Brenner, Robert Zweigerdt, Theresia Kraft
Human pluripotent stem cell (hPSC)-derived cardiomyocytes hold great potential for in vitro modeling of diseases like cardiomyopathies. Yet, knowledge about expression and functional impact of sarcomeric protein isoforms like the myosin heavy chain (MyHC) in hPSC-cardiomyocytes is scarce. We hypothesized that ventricular β-MyHC expression alters contraction and calcium kinetics and drives morphological and electrophysiological differentiation towards ventricular-like cardiomyocytes. To address this, we (1) generated human embryonic stem cell-derived cardiomyocytes (hESC-CMs) that switched towards exclusive β-MyHC, and (2) functionally and morphologically characterized these hESC-CMs at the single-cell level...
November 2016: Basic Research in Cardiology
Ilaria Piccini, Marcos Araúzo-Bravo, Guiscard Seebohm, Boris Greber
Cardiac induction of human embryonic stem cells (hESCs) is a process bearing increasing medical relevance, yet it is poorly understood from a developmental biology perspective. Anticipated technological progress in deriving stably expandable cardiac precursor cells or in advancing cardiac subtype specification protocols will likely require deeper insights into this fascinating system. Recent improvements in controlling hESC differentiation now enable a near-homogeneous induction of the cardiac lineage. This is based on an optimized initial stimulation of mesoderm-inducing signaling pathways such as Activin and/or FGF, BMP, and WNT, followed by WNT inhibition as a secondary requirement...
December 2016: Genomics Data
Wendy Keung, Lihuan Ren, Sen Li, Andy On-Tik Wong, Anant Chopra, Chi-Wing Kong, Gordon F Tomaselli, Christopher S Chen, Ronald A Li
Human embryonic stem cells (hESCs) is a potential unlimited ex vivo source of ventricular (V) cardiomyocytes (CMs), but hESC-VCMs and their engineered tissues display immature traits. In adult VCMs, sarcolemmal (sarc) and mitochondrial (mito) ATP-sensitive potassium (KATP) channels play crucial roles in excitability and cardioprotection. In this study, we aim to investigate the biological roles and use of sarcKATP and mitoKATP in hESC-VCM. We showed that SarcIK, ATP in single hESC-VCMs was dormant under baseline conditions, but became markedly activated by cyanide (CN) or the known opener P1075 with a current density that was ~8-fold smaller than adult; These effects were reversible upon washout or the addition of GLI or HMR1098...
September 28, 2016: Scientific Reports
Ruxue Wei, Xueming Zhao, Haisheng Hao, Weihua Du, Huabin Zhu
The rabbit is considered an important model animal from which to obtain embryonic stem cells because of the utility of this animal in physiology and reproductive research. Here, we derived rabbit ES-like (rES-like) cells from blastocysts of superovulated Japanese white rabbits using culture medium containing 10(-7)  M melatonin, 10 ng/mL basic fibroblast growth factor, and 1,000 IU/mL human leukemia inhibitory factor. This concentration of melatonin had the most significant positive effects on the proliferation inner cell mass-derived cells (improving rates from 19...
September 20, 2016: Molecular Reproduction and Development
Eleonora Grespan, Sebastian Martewicz, Elena Serena, Vincent Le Houérou, Jürgen Rühe, Nicola Elvassore
The mechanical activity of cardiomyocytes is the result of a process called excitation contraction coupling (ECC). A membrane depolarization wave induces a transient cytosolic calcium concentration increase that triggers activation calcium-sensitive contractile proteins leading to cell contraction and force generation. An experimental setup capable of acquiring simultaneously all ECC features would have an enormous impact on cardiac drug development and disease study. In this work, we develop a micro-engineered elastomeric substrate with tailor-made surface chemistry to measure simultaneously the uni-axial contraction force and the calcium transients generated by single human cardiomyocytes in vitro...
September 19, 2016: Langmuir: the ACS Journal of Surfaces and Colloids
Hidetoshi Masumoto, Jun K Yamashita
Stem cell therapy is a promising therapeutic option for severe cardiac diseases that are resistant to conventional therapies. To overcome the unsatisfactory results of most clinical researches on stem cell injections to an injured heart, we are developing bioengineered cardiac tissue grafts using pluripotent stem cell-derived cardiomyocytes and vascular cells. We have validated the functional benefits of mouse embryonic stem cell-derived and human induced pluripotent stem cell-derived cardiac tissue sheets (CTSs) in a rat myocardial infarction model...
November 2016: Current Treatment Options in Cardiovascular Medicine
Andre Monteiro da Rocha, Guadalupe Guerrero-Serna, Adam Helms, Carly Luzod, Sergey Mironov, Mark Russell, José Jalife, Sharlene M Day, Gary D Smith, Todd J Herron
AIMS: Mutations of cardiac sarcomere genes have been identified to cause HCM, but the molecular mechanisms that lead to cardiomyocyte hypertrophy and risk for sudden death are uncertain. The aim of this study was to examine HCM disease mechanisms at play during cardiac differentiation of human HCM specific pluripotent stem cells. METHODS AND RESULTS: We generated a human embryonic stem cell (hESC) line carrying a naturally occurring mutation of MYPBC3 (c.2905 +1 G >A) to study HCM pathogenesis during cardiac differentiation...
September 9, 2016: Journal of Molecular and Cellular Cardiology
Su Ryon Shin, Yu Shrike Zhang, Duck-Jin Kim, Ahmad Manbohi, Huseyin Avci, Antonia Silvestri, Julio Aleman, Ning Hu, Tugba Kilic, Wendy Keung, Martina Righi, Pribpandao Assawes, Hani A Alhadrami, Ronald A Li, Mehmet R Dokmeci, Ali Khademhosseini
Continual monitoring of secreted biomarkers from organ-on-a-chip models is desired to understand their responses to drug exposure in a noninvasive manner. To achieve this goal, analytical methods capable of monitoring trace amounts of secreted biomarkers are of particular interest. However, a majority of existing biosensing techniques suffer from limited sensitivity, selectivity, stability, and require large working volumes, especially when cell culture medium is involved, which usually contains a plethora of nonspecific binding proteins and interfering compounds...
October 4, 2016: Analytical Chemistry
Andrea Barbuti, Patrizia Benzoni, Giulia Campostrini, Patrizia Dell'Era
Ten years ago Yamanaka's lab identified a way to reprogram terminally differentiated cells to a pluripotent state, similar to that of embryonic stem cell. This procedure opened the road for the generation of postmitotic human cells, that have completely lost the replication potential. The initial excitement waned when it was observed that the cells produced by this method are somehow immature and do not resemble the adult phenotype. In the absence of cellular markers that recognize the various maturation steps of induced pluripotent stem cell-derived human cardiomyocytes, we propose to follow their maturation looking at their electrophysiological profile...
September 6, 2016: Developmental Dynamics: An Official Publication of the American Association of Anatomists
Jonas Schwan, Andrea T Kwaczala, Thomas J Ryan, Oscar Bartulos, Yongming Ren, Lorenzo R Sewanan, Aaron H Morris, Daniel L Jacoby, Yibing Qyang, Stuart G Campbell
We have developed an engineered heart tissue (EHT) system that uses laser-cut sheets of decellularized myocardium as scaffolds. This material enables formation of thin muscle strips whose biomechanical characteristics are easily measured and manipulated. To create EHTs, sections of porcine myocardium were laser-cut into ribbon-like shapes, decellularized, and mounted in specialized clips for seeding and culture. Scaffolds were first tested by seeding with neonatal rat ventricular myocytes. EHTs beat synchronously by day five and exhibited robust length-dependent activation by day 21...
2016: Scientific Reports
Georg Rast, Udo Kraushaar, Sandra Buckenmaier, Carina Ittrich, Brian D Guth
INTRODUCTION: Field potential duration in human pluripotent stem cell (hiPSC)-derived cardiomyocytes is discussed as parameter for the assessment of drug-induced delayed repolarization. In spontaneously beating hiPSC-derived cardiomyocytes field potential duration varies depending on beating rate but beating rate can also be influenced by field potential duration. This interdependence is not fully understood and therefore mandates careful data analysis and cautious interpretation of the results...
August 9, 2016: Journal of Pharmacological and Toxicological Methods
Anja Beckmann, Madline Schubert, Nadine Hainz, Alexandra Haase, Ulrich Martin, Thomas Tschernig, Carola Meier
Gap junction proteins are essential for direct intercellular communication but also influence cellular differentiation and migration. The expression of various connexin gap junction proteins has been demonstrated in embryonic stem cells, with Cx43 being the most intensely studied. As Cx43 is the most prominent gap junction protein in the heart, cardiomyocyte-differentiated stem cells have been studied intensely. To date, however, little is known about the expression and the subcellular distribution of Cx43 in undifferentiated stem cells or about the structural arrangement of channels...
November 2016: Histochemistry and Cell Biology
Stéphane Massé, Karl Magtibay, Nicholas Jackson, John Asta, Marjan Kusha, Boyang Zhang, Ram Balachandran, Milica Radisic, D Curtis Deno, Kumaraswamy Nanthakumar
BACKGROUND: With its inherent limitations, determining local activation times has been the basis of cardiac mapping for over a century. Here, we introduce omnipolar electrograms that originate from the natural direction of a travelling wave and from which instantaneous conduction velocity amplitude and direction can be computed at any single location without first determining activation times. We sought to validate omnipole-derived conduction velocities and explore potential application for localization of sources of arrhythmias...
July 2016: Circulation. Arrhythmia and Electrophysiology
János Pálóczi, Zoltán V Varga, Ágota Apáti, Kornélia Szebényi, Balázs Sarkadi, Rosalinda Madonna, Raffaele De Caterina, Tamás Csont, Thomas Eschenhagen, Péter Ferdinandy, Anikó Görbe
Background and Aims. Human embryonic stem cell- (hESC-) derived cardiomyocytes are one of the useful screening platforms of potential cardiocytoprotective molecules. However, little is known about the behavior of these cardiomyocytes in simulated ischemia/reperfusion conditions. In this study, we have tested the cytoprotective effect of an NO donor and the brain type natriuretic peptide (BNP) in a screening platform based first on differentiated embryonic bodies (EBs, 6 + 4 days) and then on more differentiated cardiomyocytes (6 + 24 days), both derived from hESCs...
2016: Oxidative Medicine and Cellular Longevity
Martin Pesl, Jan Pribyl, Ivana Acimovic, Aleksandra Vilotic, Sarka Jelinkova, Anton Salykin, Alain Lacampagne, Petr Dvorak, Albano C Meli, Petr Skladal, Vladimir Rotrekl
Cardiomyocyte contraction and relaxation are important parameters of cardiac function altered in many heart pathologies. Biosensing of these parameters represents an important tool in drug development and disease modeling. Human embryonic stem cells and especially patient specific induced pluripotent stem cell-derived cardiomyocytes are well established as cardiac disease model.. Here, a live stem cell derived embryoid body (EB) based cardiac cell syncytium served as a biorecognition element coupled to the microcantilever probe from atomic force microscope thus providing reliable micromechanical cellular biosensor suitable for whole-day testing...
November 15, 2016: Biosensors & Bioelectronics
Stephane Bedut, Christine Seminatore-Nole, Veronique Lamamy, Sarah Caignard, Jean A Boutin, Olivier Nosjean, Jean-Philippe Stephan, Francis Coge
Cardiomyocytes derived from human embryonic stem cells (hESCs) or induced pluripotent stem cells (hiPSCs) are increasingly used for in vitro assays and represent an interesting opportunity to increase the data throughput for drug development. In this work, we describe a 96-well recording of synchronous electrical activities from spontaneously beating hiPSC-derived cardiomyocyte monolayers. The signal was obtained with a fast-imaging plate reader using a submillisecond-responding membrane potential recording assay, FluoVolt, based on a newly derived voltage-sensitive fluorescent dye...
July 1, 2016: American Journal of Physiology. Heart and Circulatory Physiology
Fernando López-Redondo, Junko Kurokawa, Fumimasa Nomura, Tomoyuki Kaneko, Tomoyo Hamada, Tetsushi Furukawa, Kenji Yasuda
We investigated electrophysiological properties of human induced-pluripotent-stem-cell-derived and embryonic-stem-cell-derived cardiomyocytes, and analyzed action potential parameters by plotting their frequency distributions. In the both cell lines, the distribution analysis revealed that histograms of maximum upstroke velocity showed two subpopulations with similar intersection values. Sub-populations with faster maximum upstroke velocity showed significant prolongation of action potential durations by application of E-4031, whereas others did not, which may be partly due to shallower maximum diastolic potentials...
June 2016: Journal of Pharmacological Sciences
Elena Garreta, Lorena de Oñate, M Eugenia Fernández-Santos, Roger Oria, Carolina Tarantino, Andreu M Climent, Andrés Marco, Mireia Samitier, Elena Martínez, Maria Valls-Margarit, Rafael Matesanz, Doris A Taylor, Francisco Fernández-Avilés, Juan Carlos Izpisua Belmonte, Nuria Montserrat
Genome editing on human pluripotent stem cells (hPSCs) together with the development of protocols for organ decellularization opens the door to the generation of autologous bioartificial hearts. Here we sought to generate for the first time a fluorescent reporter human embryonic stem cell (hESC) line by means of Transcription activator-like effector nucleases (TALENs) to efficiently produce cardiomyocyte-like cells (CLCs) from hPSCs and repopulate decellularized human heart ventricles for heart engineering...
April 26, 2016: Biomaterials
Stephanie M Ravenscroft, Amy Pointon, Awel W Williams, Michael J Cross, James E Sidaway
The immature phenotype of stem cell derived cardiomyocytes is a significant barrier to their use in translational medicine and pre-clinical in vitro drug toxicity and pharmacological analysis. Here we have assessed the contribution of non-myocyte cells on the contractile function of co-cultured human embryonic stem cell derived cardiomyocytes (hESC-CMs) in spheroid microtissue format. Microtissues were formed using a scaffold free 96-well cell suspension method from hESC-CM cultured alone (CM microtissues) or in combination with human primary cardiac microvascular endothelial cells and cardiac fibroblasts (CMEF microtissues)...
July 2016: Toxicological Sciences: An Official Journal of the Society of Toxicology
Seong Woo Choi, Hyang-Ae Lee, Sung-Hwan Moon, Soon-Jung Park, Hae Jin Kim, Ki-Suk Kim, Yin Hua Zhang, Jae Boum Youm, Sung Joon Kim
No abstract text is available yet for this article.
April 28, 2016: Pflügers Archiv: European Journal of Physiology
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