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Cardiomyocyte Cell Culture

Monika Kasztura, Magdalena Dzięgała, Kamil Kobak, Jacek Bania, Grzegorz Mazur, Waldemar Banasiak, Piotr Ponikowski, Ewa Anita Jankowska
: Background Iron is presumed to play an important role in functioning of cardiomyocytes and skeletal myocytes. There is scarcity of direct data characterizing the cells functioning when exposed to iron depletion or iron overload in cellular environment. There is some clinical evidence demonstrating that iron deficiency has serious negative prognostic consequence in heart failure (HF) patients and its correction brought clinical benefit. BACKGROUND: Iron is presumed to play an important role in functioning of cardiomyocytes and skeletal myocytes...
October 17, 2016: Kardiologia Polska
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
Qunjun Duan, Weijun Yang, Daming Jiang, Kaiyu Tao, Aiqiang Dong, Haifeng Cheng
Myocardial infarction could result in high morbidity and mortality and heart diseases of children have becoming prevalent. Functions of spermine administration on cardiomyocytes remain unknown. The present study was designed to investigate the role of spermine pretreatment on myocardial ischemia/reperfusion injury (IRI). A cell model of simulated ischemia/reperfusion injury was established by incubating neonatal Sprague-Dawley rat cardiomyocytes in ischemia medium and re-cultured in normal medium. Of note, spermine pretreatment significantly reduced apoptosis and increased viability of immature cardiomyocytes...
2016: American Journal of Translational Research
Christopher P Jackman, Aaron L Carlson, Nenad Bursac
Engineered cardiac tissues hold promise for cell therapy and drug development, but exhibit inadequate function and maturity. In this study, we sought to significantly improve the function and maturation of rat and human engineered cardiac tissues. We developed dynamic, free-floating culture conditions for engineering "cardiobundles", 3-dimensional cylindrical tissues made from neonatal rat cardiomyocytes or human pluripotent stem cell-derived cardiomyocytes (hPSC-CMs) embedded in fibrin-based hydrogel. Compared to static culture, 2-week dynamic culture of neonatal rat cardiobundles significantly increased expression of sarcomeric proteins, cardiomyocyte size (∼2...
September 30, 2016: Biomaterials
Sean D Boothe, Jackson D Myers, Seokwon Pok, Junping Sun, Yutao Xi, Raymond M Nieto, Jie Cheng, Jeffrey G Jacot
The stiffness of myocardial tissue changes significantly at birth and during neonatal development, concurrent with significant changes in contractile and electrical maturation of cardiomyocytes. Previous studies by our group have shown that cardiomyocytes generate maximum contractile force when cultured on a substrate with a stiffness approximating native cardiac tissue. However, effects of substrate stiffness on the electrophysiology and ion currents in cardiomyocytes have not been fully characterized. In this study, neonatal rat ventricular myocytes were cultured on the surface of flat polyacrylamide hydrogels with elastic moduli ranging from 1 to 25 kPa...
October 8, 2016: Cell Biochemistry and Biophysics
Hai-Feng Pei, Juan-Ni Hou, Fei-Peng Wei, Qiang Xue, Fan Zhang, Cheng-Fei Peng, Yi Yang, Yue Tian, Juan Feng, Jin Du, Lei He, Xiu-Chuan Li, Er-He Gao, De Li, Yong-Jian Yang
Mitochondrial dysfunction leads to reactive oxygen species (ROS) overload, exacerbating injury in myocardial infarction (MI). As a receptor for translocases in the outer mitochondrial membrane (Tom) complex, Tom70 has an unknown function in MI, including in melatonin-induced protection against MI injury. We delivered specific small interfering RNAs against Tom70 or lentivirus vectors carrying Tom70a sequences into the left ventricles of mice or to cultured neonatal murine ventricular myocytes (NMVMs). At 48 h post-transfection, the left anterior descending coronary arteries of mice were permanently ligated, while the NMVMs underwent continuous hypoxia...
October 5, 2016: Journal of Pineal Research
M Borrell-Pages, G Vilahur, J C Romero, L Casaní, M T Bejar, L Badimon
LRP5 (low-density lipoprotein receptor-related protein 5) activates canonical Wnt signalling. LRP5 plays multiple roles including regulation of lipoprotein and cholesterol homeostasis as well as innate immunity cell function. However, it is not known whether LRP5 has a role in the myocardium. The aim of this study was to investigate LRP5 and Wnt signalling in myocardial remodelling after acute myocardial infarction (MI). Wnt protein levels were determined in a hypercholesterolemic porcine model of MI, in Lrp5 (-/-) C57Bl6 mice, in cultured cardiomyocytes and in human explanted hearts with previous MI episodes...
November 2016: Basic Research in Cardiology
Brett J Roberts, Robert A Svoboda, Andrew M Overmiller, Joshua D Lewis, Andrew P Kowalczyk, My G Mahoney, Keith R Johnson, James K Wahl
Desmosomes are prominent adhesive junctions present between many epithelial cells as well as cardiomyocytes. The mechanisms controlling desmosome assembly and remodeling in epithelial and cardiac tissue are poorly understood. We recently identified protein palmitoylation as a mechanism regulating desmosome dynamics. In this study we have focused on the palmitoylation of the desmosomal cadherin, desmoglein-2 (Dsg2), and characterized the role that palmitoylation of Dsg2 plays in its localization and stability in cultured cells...
October 4, 2016: Journal of Biological Chemistry
Ming-Ming Xi, Jing Liu, Liang Xie, Jian-Bin Gong
OBJECTIVE: To investigated the effects of bisoprolol pretreatment on hypoxia/reoxygenation (H/R)-induced injury in H9c2 cardiomyocytes. METHODS: Cultured H9c2 cells were exposed to hypoxia for 6 h followed by reoxygenation for 2 h with or without pretreatments with bisoprolol or bisoprolol + LY294002. The cell survival was measured by MTT assay, and the cell apoptosis and levels of radical oxygen species (ROS) were evaluated with flow cytometry. The protein levels of phosphyorylated AKT and phosphorylated GSK3β in the cells were determined by Western blotting...
August 20, 2016: Nan Fang Yi Ke da Xue Xue Bao, Journal of Southern Medical University
Justin Judd, Jonathan Lovas, Guo N Huang
Cultured cardiomyocytes can be used to study cardiomyocyte biology using techniques that are complementary to in vivo systems. For example, the purity and accessibility of in vitro culture enables fine control over biochemical analyses, live imaging, and electrophysiology. Long-term culture of cardiomyocytes offers access to additional experimental approaches that cannot be completed in short term cultures. For example, the in vitro investigation of dedifferentiation, cell cycle re-entry, and cell division has thus far largely been restricted to rat cardiomyocytes, which appear to be more robust in long-term culture...
2016: Journal of Visualized Experiments: JoVE
Jin Yu, Yiliyaer Maimaitili, Peng Xie, Jianjiang Wu, Jiang Wang, Yining Yang, Haiping Ma, Hong Zheng
AIM: Hyperglycaemia-induced cell injury is a primary cause of cardiovascular complications in diabetic patients. In vivo studies demonstrated that sevoflurane postconditioning (SpostC) was cardioprotective against ischaemia/reperfusion injury, which was blocked by hyperglycaemia. This study investigated whether high glucose concentration abrogated SpostC cardioprotection in vitro by advancing mitochondrial fission and whether mitochondrial division inhibitor-1 (Mdivi-1) restored SpostC cardioprotection in cultured primary neonatal rat cardiomyocytes (NCMs)...
September 29, 2016: Acta Physiologica
Ahmet U Uzun, Ingra Mannhardt, Kaja Breckwoldt, András Horváth, Silke S Johannsen, Arne Hansen, Thomas Eschenhagen, Torsten Christ
Human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CM) provide a unique opportunity to study human heart physiology and pharmacology and repair injured hearts. The suitability of hiPSC-CM critically depends on how closely they share physiological properties of human adult cardiomyocytes (CM). Here we investigated whether a 3D engineered heart tissue (EHT) culture format favors maturation and addressed the L-type Ca(2+)-current (ICa,L) as a readout. The results were compared with hiPSC-CM cultured in conventional monolayer (ML) and to our previous data from human adult atrial and ventricular CM obtained when identical patch-clamp protocols were used...
2016: Frontiers in Pharmacology
Martin D Brand, Renata L S Goncalves, Adam L Orr, Leonardo Vargas, Akos A Gerencser, Martin Borch Jensen, Yves T Wang, Simon Melov, Carolina N Turk, Jason T Matzen, Victoria J Dardov, H Michael Petrassi, Shelly L Meeusen, Irina V Perevoshchikova, Heinrich Jasper, Paul S Brookes, Edward K Ainscow
Using high-throughput screening we identified small molecules that suppress superoxide and/or H2O2 production during reverse electron transport through mitochondrial respiratory complex I (site IQ) without affecting oxidative phosphorylation (suppressors of site IQ electron leak, "S1QELs"). S1QELs diminished endogenous oxidative damage in primary astrocytes cultured at ambient or low oxygen tension, showing that site IQ is a normal contributor to mitochondrial superoxide-H2O2 production in cells. They diminished stem cell hyperplasia in Drosophila intestine in vivo and caspase activation in a cardiomyocyte cell model driven by endoplasmic reticulum stress, showing that superoxide-H2O2 production by site IQ is involved in cellular stress signaling...
October 11, 2016: Cell Metabolism
Marcela Pizarro, Rodrigo Troncoso, Gonzalo J Martínez, Mario Chiong, Pablo F Castro, Sergio Lavandero
Doxorubicin (Doxo) is one of the most effective anti-neoplastic agents but its cardiotoxicity has been an important clinical limitation. The major mechanism of Doxo-induced cardiotoxicity is associated to its oxidative capacity. However, other processes are also involved with significant consequences for the cardiomyocyte. In recent years, a number of studies have investigated the role of autophagy on Doxo-induced cardiotoxicity but to date it is not clear how Doxo alters that process and its consequence on cardiomyocytes viability...
September 22, 2016: Toxicology
Elena Dyukova, Rolf Schreckenberg, Christoph Arens, Guzel Sitdikova, Klaus-Dieter Schlüter
Nitric oxide (NO)-deficiency as it occurs during endothelial dysfunction activates the endothelin-1 system and increases the expression of receptor activity modifying protein (RAMP)-1 that acts as a chaperon for calcium-sensing receptors (CaR) that have recently been identified to improve cardiac function. Here we hypothesized that endothelin-1 increases the cardiac expression of CaR and thereby induces an adaptive type of hypertrophy. Expressions of RAMP-1, endothelin receptors, and CaR were analyzed by RT-PCR in left ventricular tissues of L-NAME-treated rats...
September 23, 2016: Journal of Cellular Physiology
Ellis Y Kim, Patrick Page, Lisa M Dellefave-Castillo, Elizabeth M McNally, Eugene J Wyatt
BACKGROUND: Cellular models of muscle disease are taking on increasing importance with the large number of genes and mutations implicated in causing myopathies and the concomitant need to test personalized therapies. Developing cell models relies on having an easily obtained source of cells, and if the cells are not derived from muscle itself, a robust reprogramming process is needed. Fibroblasts are a human cell source that works well for the generation of induced pluripotent stem cells, which can then be differentiated into cardiomyocyte lineages, and with less efficiency, skeletal muscle-like lineages...
2016: Skeletal Muscle
Wei Yang, Zhaoya Liu, Qian Xu, Haiyang Peng, Luyao Chen, Xiao Huang, Tianlun Yang, Zaixin Yu, Guangjie Cheng, Guogang Zhang, Ruizheng Shi
Oxidative stress has been implicated in cardiac hypertrophy and heart failure. Vascular peroxidase 1 (VPO1), a peroxidase in the cardiovascular system, uses the hydrogen peroxide (H2O2) derived from co-expressed NADPH oxidases (NOX) to produce hypochlorous acid (HOCl) and catalyze peroxidative reactions. Our previous studies showed that VPO1 contributes to the vascular smooth muscle cell proliferation and endothelial dysfunction in spontaneous hypertensive rats (SHRs); however, the role of VPO1 in cardiomyocytes hypertrophy is still uninvestigated...
August 24, 2016: Journal of the American Society of Hypertension: JASH
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
Luca Sala, Milena Bellin, Christine L Mummery
Cardiotoxicity is a severe side effect of drugs that induce structural or electrophysiological changes in heart muscle cells. As a result, the heart undergoes failure and potentially lethal arrhythmias. It is still a major reason for drug failure in preclinical and clinical phases of drug discovery. Current methods for predicting cardiotoxicity are based on guidelines which combine electrophysiological analysis of cell lines expressing ion channels ectopically in vitro with animal models and clinical trials...
September 19, 2016: British Journal of Pharmacology
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