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Cardiomyocyte Cell Culture

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https://www.readbyqxmd.com/read/28813698/caspase-independent-pathway-is-related-to-nilotinib-cytotoxicity-in-cultured-cardiomyocytes
#1
Qinghui Yang, Chunhui Zhang, Hong Wei, Zenghui Meng, Guangnan Li, Yuanyuan Xu, Yanjun Chen
BACKGROUND/AIMS: Cardiotoxicity is a predominant side-effect of nilotinib during chronic myeloid leukemia treatment. The underlying molecular mechanism remains unclear. The role of autophagy and mitochondrial signaling was investigated in nilotinib-treated cardiac H9C2 cells. METHODS: Cytotoxicity was assessed using Cell Death Detection kit. Immunoblot and immunofluorescence staining was performed, and cathepsin B and caspase3 activity was assessed in nilotinib-treated H9C2 cells with or without distinct pathway inhibitor or specific siRNA...
August 15, 2017: Cellular Physiology and Biochemistry
https://www.readbyqxmd.com/read/28808223/metastasis-associated-protein-1-deficiency-results-in-compromised-pulmonary-alveolar-capillary-angiogenesis-in-mice
#2
Jun-Hui Qin, Zhen-Yu Ke, Qiang Zhou, Li Wang, Yuan Liang, Ying-Mei Wang, Tong Yang, Xing Gao, Jing Ye, Rekesh Kumar, Rui-An Wang
BACKGROUND The aim of this study was to investigate the effects of metastasis-associated protein 1 (MTA1) deficiency during angiogenesis of pulmonary alveolar capillaries in mice and to determine the molecular mechanisms involved. MATERIAL AND METHODS The expressions of MTA1, CD34, vascular endothelial growth factor (VEGF), alpha smooth muscle actin (α-SMA), and HIF-1α were analyzed in the lungs of MTA1-knockout (KO) and wild-type mice at embryonic day 18.5 and 2 months by quantitative PCR, immunoblotting, and immunohistochemistry...
August 15, 2017: Medical Science Monitor: International Medical Journal of Experimental and Clinical Research
https://www.readbyqxmd.com/read/28806493/olaparib-a-clinically-used-poly-adp-ribose-polymerase-inhibitor-protects-against-oxidant-induced-cardiac-myocyte-death-in-vitro-and-improves-cardiac-contractility-during-early-phase-after-heart-transplantation-in-a-rat-model-in-vivo
#3
Sevil Korkmaz-Icöz, Bartosz Szczesny, Michela Marcatti, Shiliang Li, Mihály Ruppert, Felix Lasitschka, Sivakkanan Loganathan, Csaba Szabo, Gábor Szabó
BACKGROUND AND PURPOSE: Olaparib, rucaparib and niraparib, potent inhibitors of poly(ADP-ribose) polymerase (PARP) have recently been approved for human use for oncological indications. Considering the previously demonstrated role of PARP in various forms of acute and chronic myocardial injury, we tested the effect of olaparib in in-vitro models of oxidative stress in cardiomyocytes, and in an in-vivo model of cardiac transplantation. EXPERIMENTAL APPROACH: H9c2-embryonic rat heart-derived myoblasts pretreated with vehicle or olaparib (10μM) were challenged with either hydrogen-peroxide (H2 O2 ) or with glucose-oxidase (GOx, which generates H2 O2 in the tissue culture medium)...
August 14, 2017: British Journal of Pharmacology
https://www.readbyqxmd.com/read/28801548/virtual-cardiac-monolayers-for-electrical-wave-propagation
#4
Nina Kudryashova, Valeriya Tsvelaya, Konstantin Agladze, Alexander Panfilov
The complex structure of cardiac tissue is considered to be one of the main determinants of an arrhythmogenic substrate. This study is aimed at developing the first mathematical model to describe the formation of cardiac tissue, using a joint in silico-in vitro approach. First, we performed experiments under various conditions to carefully characterise the morphology of cardiac tissue in a culture of neonatal rat ventricular cells. We considered two cell types, namely, cardiomyocytes and fibroblasts. Next, we proposed a mathematical model, based on the Glazier-Graner-Hogeweg model, which is widely used in tissue growth studies...
August 11, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28801517/effect-of-densely-ionizing-radiation-on-cardiomyocyte-differentiation-from-human-induced-pluripotent-stem-cells
#5
Erdene Baljinnyam, Sundararajan Venkatesh, Richard Gordan, Satvik Mareedu, Jianyi Zhang, Lai-Hua Xie, Edouard I Azzam, Carolyn K Suzuki, Diego Fraidenraich
The process of human cardiac development can be faithfully recapitulated in a culture dish with human pluripotent stem cells, where the impact of environmental stressors can be evaluated. The consequences of ionizing radiation exposure on human cardiac differentiation are largely unknown. In this study, human-induced pluripotent stem cell cultures (hiPSCs) were subjected to an external beam of 3.7 MeV α-particles at low mean absorbed doses of 0.5, 3, and 10 cGy. Subsequently, the hiPSCs were differentiated into beating cardiac myocytes (hiPSC-CMs)...
August 2017: Physiological Reports
https://www.readbyqxmd.com/read/28791052/inhibition-of-histone-methyltransferase-histone-deacetylase-and-%C3%AE-catenin-synergistically-enhance-the-cardiac-potential-of-bone-marrow-cells
#6
Jinpu Yang, Keerat Kaur, John G Edwards, Carol A Eisenberg, Leonard M Eisenberg
Previously, we reported that treatment with the G9a histone methyltransferase inhibitor BIX01294 causes bone marrow mesenchymal stem cells (MSCs) to exhibit a cardiocompetent phenotype, as indicated by the induction of the precardiac markers Mesp1 and brachyury. Here, we report that combining the histone deacetylase inhibitor trichostatin A (TSA) with BIX01294 synergistically enhances MSC cardiogenesis. Although TSA by itself had no effect on cardiac gene expression, coaddition of TSA to MSC cultures enhanced BIX01294-induced levels of Mesp1 and brachyury expression 5...
2017: Stem Cells International
https://www.readbyqxmd.com/read/28771828/vegfb-vegfr1-ameliorates-ang-ii-induced-cardiomyocyte-hypertrophy-through-ca-2-mediated-pkg-i-pathway
#7
Zhijie Shen, Zhihui Zhang, Xiaoyan Wang, Kan Yang
In response to assorted stimuli, the heart will develop into cardiomyocyte hypertrophy, but sustained cardiomyocyte hypertrophy will finally lead to heart failure. This research is aimed to examine the effect of VEGFB on cardiomyocyte hypertrophy by using the cardiomyocyte-derived cell line H9C2 of cultured rates. It turns out that VEGFB can positively prevent the Ang II-induced rising in the size of cardiomyocyte as well as reduce Ang II-induced mRNA and protein levels of β-MHC (β-myosin heavy chain), BNP (brain natriuretic peptide) and ANP (atrial natriuretic peptide)...
August 3, 2017: Journal of Cellular Biochemistry
https://www.readbyqxmd.com/read/28765558/cardiac-spheroids-as-promising-in-vitro-models-to-study-the-human-heart-microenvironment
#8
Liudmila Polonchuk, Mamta Chabria, Laura Badi, Jean-Christophe Hoflack, Gemma Figtree, Michael J Davies, Carmine Gentile
Three-dimensional in vitro cell systems are a promising alternative to animals to study cardiac biology and disease. We have generated three-dimensional in vitro models of the human heart ("cardiac spheroids", CSs) by co-culturing human primary or iPSC-derived cardiomyocytes, endothelial cells and fibroblasts at ratios approximating those present in vivo. The cellular organisation, extracellular matrix and microvascular network mimic human heart tissue. These spheroids have been employed to investigate the dose-limiting cardiotoxicity of the common anti-cancer drug doxorubicin...
August 1, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28748466/impedance-measurement-in-induced-pluripotent-stem-cell-derived-cardiomyocytes
#9
Emma-Jane Poulton
Impedance-based measurement is a useful tool for assessing the contractility of plated cardiomyocytes in the context of early preclinical cardiosafety assessment. Induced pluripotent stem cell-derived cardiomyocytes (iPSCs) can be used for this purpose as these cells display similar electrochemical properties to primary cardiomyocytes and beat reliably and in synchronicity in culture. Here we describe a method for measuring the contractility of iPSCs using the xCELLigence RTCA impedance measurement system.
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28747015/nardilysin-is-a-promising-biomarker-for-the-early-diagnosis-of-acute-coronary-syndrome
#10
Po-Min Chen, Mikiko Ohno, Takaki Hiwasa, Kiyoto Nishi, Sayaka Saijo, Jiro Sakamoto, Yusuke Morita, Shintaro Matsuda, Shin Watanabe, Yasuhide Kuwabara, Koh Ono, Masao Imai, Katsumi Inoue, Tatsuya Murai, Tsukasa Inada, Masaru Tanaka, Toru Kita, Takeshi Kimura, Eiichiro Nishi
BACKGROUND: Biomarkers for detection of transient myocardial ischemia in patients with unstable angina (UA) or for very early diagnosis of acute myocardial infarction (AMI) are not currently available. METHODS AND RESULTS: We performed two sequential screenings of autoantibodies elevated shortly after the onset of acute coronary syndrome (ACS), and focused on metalloendopeptidase nardilysin (NRDC) among 19 identified candidate antigens. In a retrospective analysis among 93 ACS and 117 non-ACS patients, the serum level of NRDC was significantly increased in patients with ACS compared with that in patients with non-ACS (2073...
September 15, 2017: International Journal of Cardiology
https://www.readbyqxmd.com/read/28742197/mir-24-alleviates-cardiomyocyte-apoptosis-after-myocardial-infarction-via-targeting-bim
#11
L-J Pan, X Wang, Y Ling, H Gong
OBJECTIVE: Ischemia hypoxia induces cardiomyocyte (CM) apoptosis in the process of acute myocardial infarction (AMI). It was showed that pro-apoptosis factor BIM participates in regulating tumor cell apoptosis under ischemia or hypoxia condition, while its role in CM apoptosis after AMI is still unclear. It was revealed that miR-24 expression was significantly reduced in myocardial tissue after AMI. Bioinformatics analysis exhibits that miR-24 is targeted to the 3'-UTR of BIM. This study aims to investigate the role of miR-24 in mediating BIM expression and CM apoptosis...
July 2017: European Review for Medical and Pharmacological Sciences
https://www.readbyqxmd.com/read/28735524/characterization-of-cytoskeleton-features-and-maturation-status-of-cultured-human-ipsc-derived-cardiomyocytes
#12
Christian Zuppinger, George Gibbons, Priyanka Dutta-Passecker, Adrian Segiser, Henriette Most, Thomas M Suter
Recent innovations in stem cell technologies and the availability of human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) have opened new possibilities for studies and drug testing on human cardiomyocytes in vitro. Still, there are concerns about the precise nature of such 'reprogrammed' cells. We have performed an investigation using immunocytochemistry and confocal microscopy on several cellular features using commercially available hiPSC-CMs. For some selected developmentally regulated or cardiac chamber-specific proteins, we have compared the results from hiPSC-derived cardiomyocytes with freshly isolated, ventricular cardiomyocytes from adult rats...
June 21, 2017: European Journal of Histochemistry: EJH
https://www.readbyqxmd.com/read/28734787/selenium-deficiency-induced-thioredoxin-suppression-and-thioredoxin-knock-down-disbalanced-insulin-responsiveness-in-chicken-cardiomyocytes-through-pi3k-akt-pathway-inhibition
#13
Jie Yang, Sattar Hamid, Jingzeng Cai, Qi Liu, Shiwen Xu, Ziwei Zhang
Thioredoxin (Txn) system is the most crucial antioxidant defense mechanism in cell consisting of Txn, thioredoxin reductase (TR) and Nicotinamide Adenine Dinucleotide Phosphate (NADPH). Perturbations in Txn system may compromise cell survival through oxidative stress induction. Metabolic activity of insulin plays important roles in fulfilling the stable and persistent demands of heart through glucose metabolism. However, the roles of Txn and Txn system in insulin modulated cardiac energy metabolism have been less reported...
July 19, 2017: Cellular Signalling
https://www.readbyqxmd.com/read/28731525/differences-in-stem-cell-processing-lead-to-distinct-secretomes-secretion-implications-for-differential-results-of-previous-clinical-trials-of-stem-cell-therapy-for-myocardial-infarction
#14
Bernhard Wernly, Inês Gonçalves, Attila Kiss, Vera Paar, Tobias Mösenlechner, Michael Leisch, David Santer, Lucas Motloch, Klaus Ulrich Klein, Eva Verena Tretter, Daniel Kretzschmar, Bruno Podesser, Christian Jung, Uta C Hoppe, Michael Lichtenauer
INTRODUCTION: Stem cell therapy for acute myocardial infarction (AMI) seemed to be a promising therapy, however large clinical trials brought differential outcome. It has been shown that paracrine effects of secretomes of stem cells rather than cell therapy might play a fundamental role. The present study sought to compare cell processing protocols of clinical trials and investigated effects of differential cell culture conditions on chemokine secretion and functional effects. METHODS: Different secretomes were compared regarding IL-8, VEGF, MCP-1 and TNF-alpha secretion...
July 21, 2017: Biotechnology Journal
https://www.readbyqxmd.com/read/28729840/cardiac-subtype-specific-modeling-of-kv1-5-ion-channel-deficiency-using-human-pluripotent-stem-cells
#15
Maike Marczenke, Ilaria Piccini, Isabella Mengarelli, Jakob Fell, Albrecht Röpke, Guiscard Seebohm, Arie O Verkerk, Boris Greber
The ultrarapid delayed rectifier K(+) current (IKur), mediated by Kv1.5 channels, constitutes a key component of the atrial action potential. Functional mutations in the underlying KCNA5 gene have been shown to cause hereditary forms of atrial fibrillation (AF). Here, we combine targeted genetic engineering with cardiac subtype-specific differentiation of human induced pluripotent stem cells (hiPSCs) to explore the role of Kv1.5 in atrial hiPSC-cardiomyocytes. CRISPR/Cas9-mediated mutagenesis of integration-free hiPSCs was employed to generate a functional KCNA5 knockout...
2017: Frontiers in Physiology
https://www.readbyqxmd.com/read/28727799/atlantic-salmon-cardiac-primary-cultures-an-in-vitro-model-to-study-viral-host-pathogen-interactions-and-pathogenesis
#16
Patricia A Noguera, Bianka Grunow, Matthias Klinger, Katherine Lester, Bertrand Collet, Jorge Del-Pozo
Development of Salmon Cardiac Primary Cultures (SCPCs) from Atlantic salmon pre-hatch embryos and their application as in vitro model for cardiotropic viral infection research are described. Producing SCPCs requires plating of trypsin dissociated embryos with subsequent targeted harvest from 24h up to 3 weeks, of relevant tissues after visual identification. SCPCs are then transferred individually to chambered wells for culture in isolation, with incubation at 15-22°. SCPCs production efficiency was not influenced by embryo's origin (0...
2017: PloS One
https://www.readbyqxmd.com/read/28727019/triennial-growth-and-development-symposium-dedifferentiated-fat-cells-potential-and-perspectives-for-their-use-in-clinical-and-animal-science-purpose
#17
M S Duarte, R Bueno, W Silva, C F Campos, M P Gionbelli, S E F Guimarães, F F Silva, P S Lopes, G J Hausman, M V Dodson
An increasing body of evidences has demonstrated the ability of the mature adipocyte to dedifferentiate into a population of proliferative-competent cells known as dedifferentiated fat (DFAT) cells. As early as the 1970s, in vitro studies showed that DFAT cells may be obtained by ceiling culture, which takes advantage of the buoyancy property of lipid-filled cells. It was documented that DFAT cells may acquire a phenotype similar to mesenchymal stem cells and yet may differentiate into multiple cell lineages, such as skeletal and smooth muscle cells, cardiomyocytes, osteoblasts, and adipocytes...
May 2017: Journal of Animal Science
https://www.readbyqxmd.com/read/28715970/scaffold-in-scaffold-potential-to-induce-growth-and-differentiation-of-cardiac-progenitor-cells
#18
Matteo Ciocci, Federico Mochi, Felicia Carotenuto, Emilia Di Giovanni, Paolo Prosposito, Roberto Francini, Fabio De Matteis, Igor Reshetov, Mauro Casalboni, Sonia Melino, Paolo Di Nardo
The design of reliable biocompatible and biodegradable scaffolds remains one of the most important challenges for tissue engineering. In fact, properly designed scaffolds must display an adequate and interconnected porosity to facilitate cell spreading and colonization of the inner layers, and must release physical signals concurring to modulate cell function to ultimately drive cell fate. In the present study, a combination of optimal mechanical and biochemical properties has been considered to design a one-component 3D multitextured hydrogel scaffold in order to favor cell-scaffold interactions...
July 17, 2017: Stem Cells and Development
https://www.readbyqxmd.com/read/28715377/creation-of-cardiac-tissue-exhibiting-mechanical-integration-of-spheroids-using-3d-bioprinting
#19
Chin Siang Ong, Takuma Fukunishi, Andrew Nashed, Adriana Blazeski, Huaitao Zhang, Samantha Hardy, Deborah DiSilvestre, Luca Vricella, John Conte, Leslie Tung, Gordon Tomaselli, Narutoshi Hibino
This protocol describes 3D bioprinting of cardiac tissue without the use of biomaterials, using only cells. Cardiomyocytes, endothelial cells and fibroblasts are first isolated, counted and mixed at desired cell ratios. They are co-cultured in individual wells in ultra-low attachment 96-well plates. Within 3 days, beating spheroids form. These spheroids are then picked up by a nozzle using vacuum suction and assembled on a needle array using a 3D bioprinter. The spheroids are then allowed to fuse on the needle array...
July 2, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28715029/anisotropic-microfibrous-scaffolds-enhance-the-organization-and-function-of-cardiomyocytes-derived-from-induced-pluripotent-stem-cells
#20
Maureen Wanjare, Luqia Hou, Karina H Nakayama, Joseph J Kim, Nicholas P Mezak, Oscar J Abilez, Evangeline Tzatzalos, Joseph C Wu, Ngan F Huang
Engineering of myocardial tissue constructs is a promising approach for treatment of coronary heart disease. To engineer myocardial tissues that better mimic the highly ordered physiological arrangement and function of native cardiomyocytes, we generated electrospun microfibrous polycaprolactone scaffolds with either randomly oriented (14 μm fiber diameter) or parallel-aligned (7 μm fiber diameter) microfiber arrangement and co-seeded the scaffolds with human induced pluripotent stem cell-derived cardiomyocytes (iCMs) and endothelial cells (iECs) for up to 12 days after iCM seeding...
July 25, 2017: Biomaterials Science
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