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Cardiomyocyte Cell Culture

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https://www.readbyqxmd.com/read/28343439/toward-improved-myocardial-maturity-in-an-organ-on-chip-platform-with-immature-cardiac-myocytes
#1
Sean P Sheehy, Anna Grosberg, Pu Qin, David J Behm, John P Ferrier, Mackenzie A Eagleson, Alexander P Nesmith, David Krull, James G Falls, Patrick H Campbell, Megan L McCain, Robert N Willette, Erding Hu, Kevin K Parker
In vitro studies of cardiac physiology and drug response have traditionally been performed on individual isolated cardiomyocytes or isotropic monolayers of cells that may not mimic desired physiological traits of the laminar adult myocardium. Recent studies have reported a number of advances to Heart-on-a-Chip platforms for the fabrication of more sophisticated engineered myocardium, but cardiomyocyte immaturity remains a challenge. In the anisotropic musculature of the heart, interactions between cardiac myocytes, the extracellular matrix (ECM), and neighboring cells give rise to changes in cell shape and tissue architecture that have been implicated in both development and disease...
January 1, 2017: Experimental Biology and Medicine
https://www.readbyqxmd.com/read/28337264/ip3r-and-ryr-calcium-channels-are-involved-in-neonatal-rat-cardiac-myocyte-hypertrophy-induced-by-tumor-necrosis-factor-%C3%AE
#2
Gui-Jun Wang, Lian-Yi Guo, Hong-Xin Wang, Yu-Sheng Yao
To investigate which calcium channels are involved in cardiac myocyte hypertrophy induced by TNF-α, cultured cardiomyocytes were treated with 100 μg/L TNF-α. In addition, three different calcium channel blockers (2-APB, ryanodine and nifedipine) were used, and the effects of each calcium channel blocker on cardiac hypertrophy induced by TNF-α were carefully observed. Measurements included cytosolic calcium transients ([Ca(2+)]i), the level of intracellular calcium in individual cells, cell protein content, cell protein synthesis and cell volume...
2017: American Journal of Translational Research
https://www.readbyqxmd.com/read/28336914/inhibition-of-serum-and-glucocorticoid-regulated-kinase-1-as-novel-therapy-for-cardiac-arrhythmia-disorders
#3
Vassilios J Bezzerides, Aifeng Zhang, Ling Xiao, Bridget Simonson, Santosh A Khedkar, Shiro Baba, Filomena Ottaviano, Stacey Lynch, Katherine Hessler, Alan C Rigby, David Milan, Saumya Das, Anthony Rosenzweig
Alterations in sodium flux (INa) play an important role in the pathogenesis of cardiac arrhythmias and may also contribute to the development of cardiomyopathies. We have recently demonstrated a critical role for the regulation of the voltage-gated sodium channel NaV1.5 in the heart by the serum and glucocorticoid regulated kinase-1 (SGK1). Activation of SGK1 in the heart causes a marked increase in both the peak and late sodium currents leading to prolongation of the action potential duration and an increased propensity to arrhythmia...
March 23, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28316060/hotair-functions-as-a-competing-endogenous-rna-to-regulate-pten-expression-by-inhibiting-mir-19-in-cardiac-hypertrophy
#4
Yanjun Lai, Shuai He, Liming Ma, Hong Lin, Biyun Ren, Jing Ma, Xinyu Zhu, Shifang Zhuang
Sustained cardiac hypertrophy (CH) is related to a variety of physiological as well as pathological stimuli and eventually increases the risk of heart failure. HOTAIR has been identified as a competing endogenous RNA in multiple human biological processes. Whether lncRNA-HOTAIR is involved in the progress of CH and how it works still remain unknown. Herein, we found that HOTAIR was down-regulated, while miR-19 was up-regulated in both heart tissues from TAC-operated mice in vivo and cultural cardiomyocytes treated with Ang-II in vitro by real-time PCR...
March 18, 2017: Molecular and Cellular Biochemistry
https://www.readbyqxmd.com/read/28295592/knockdown-of-long-non-coding-rna-zfas1-protects-cardiomyocytes-against-acute-myocardial-infarction-via-anti-apoptosis-by-regulating-mir-150-crp
#5
Tao Wu, Dan Wu, Qinghua Wu, Bing Zou, Xiao Huang, Xiaoshu Cheng, Yanqing Wu, Kui Hong, Ping Li, Renqiang Yang, Yunde Li, Yingzhang Cheng
BACKGROUND: ZFAS1 is one of cardiac-specific or cardiac-related lncRNAs. This study was to explore the functional involvement of ZFAS1 and its regulatory role in AMI. METHODS: In this study, the models of AMI rat and myocardial cell cultured under hypoxia were made. The expression of ZFAS1 and miR-150 of myocardial infarction tissue or cardiac myocytes was determined by quantitative real time PCR. The regulatory role of ZFAS1 on miR-150 was examined by RNA pull down assay...
March 10, 2017: Journal of Cellular Biochemistry
https://www.readbyqxmd.com/read/28288171/the-acceleration-of-cardiomyogenesis-in-embryonic-stem-cells-in-vitro-by-serum-depletion-does-not-increase-the-number-of-developed-cardiomyocytes
#6
Katarzyna Anna Radaszkiewicz, Dominika Sýkorová, Lucia Binó, Jana Kudová, Markéta Bébarová, Jiřina Procházková, Hana Kotasová, Lukáš Kubala, Jiří Pacherník
The differentiation of pluripotent embryonic stem (ES) cells into various lineages in vitro represents an important tool for studying the mechanisms underlying mammalian embryogenesis. It is a key technique in studies evaluating the molecular mechanisms of cardiomyogenesis and heart development and also in embryotoxicology. Herein, modest modifications of the basic protocol for ES cell differentiation into cardiomyocytes were evaluated in order to increase the yield and differentiation status of developed cardiomyocytes...
2017: PloS One
https://www.readbyqxmd.com/read/28283792/extracellular-vesicles-mediated-transfer-of-mir-208a-b-exaggerate-hypoxia-reoxygenation-injury-in-cardiomyocytes-by-reducing-qki-expression
#7
Feng Wang, Yuxiang Yuan, Pirong Yang, Xia Li
In this study, we tested the hypothesis that extracellular vesicles (EVs)-mediated transfer of miR-208a/b can exacerbate apoptosis of cardiomyocytes (CMs) induced by hypoxia/reoxygenation (H/R) injury by reducing the expression of the RNA-binding protein Quaking (QKI). EVs were isolated from culture medium of hypoxic H9c2 cells (EVs-H). In in vitro H9c2 cell model, the EVs-H could be taken up by normoxic CMs and exacerbated cell apoptosis induced by H/R injury. In addition, miR-208a and miR-208b were enriched in EVs-H...
March 10, 2017: Molecular and Cellular Biochemistry
https://www.readbyqxmd.com/read/28283550/the-effects-of-%C3%AE-adrenergic-receptor-drugs-on-embryonic-ventricular-cell-proliferation-and-differentiation-and-their-impact-on-donor-cell-transplantation
#8
Tiam Feridooni, Adam Hotchkiss, Mark Baguma-Nibasheka, Feixiong Zhang, Brittney Allen, Sarita Chinni, Kishore B S Pasumarthi
The beta-adrenergic receptors (β-ARs) and catecholamines are present in rodents as early as embryonic day 10.5 (E10.5). However, it is not known whether β-AR signaling plays any role in proliferation and differentiation of ventricular cells in the embryonic heart. Here, we characterized the expression profiles of β-AR subtypes and established dose response curves for a nonselective β-AR agonist isoproterenol (ISO) in the developing mouse ventricular cells. Further, we investigated the effects of ISO on cell cycle activity and differentiation of cultured E11...
March 10, 2017: American Journal of Physiology. Heart and Circulatory Physiology
https://www.readbyqxmd.com/read/28266620/two-dimensional-electrophysiological-characterization-of-human-pluripotent-stem-cell-derived-cardiomyocyte-system
#9
Huanqi Zhu, Kelsey S Scharnhorst, Adam Z Stieg, James K Gimzewski, Itsunari Minami, Norio Nakatsuji, Haruko Nakano, Atsushi Nakano
Stem cell-derived cardiomyocytes provide a promising tool for human developmental biology, regenerative therapies, disease modeling, and drug discovery. As human pluripotent stem cell-derived cardiomyocytes remain functionally fetal-type, close monitoring of electrophysiological maturation is critical for their further application to biology and translation. However, to date, electrophysiological analyses of stem cell-derived cardiomyocytes has largely been limited by biologically undefined factors including 3D nature of embryoid body, sera from animals, and the feeder cells isolated from mouse...
March 7, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28260010/astragaloside-iv-inhibits-isoprenaline%C3%A2-induced-cardiac-fibrosis-by-targeting-the-reactive-oxygen-species-mitogen%C3%A2-activated-protein-kinase-signaling-axis
#10
Hongliang Dai, Guizhi Jia, Meili Lu, Chunguang Liang, Yue Wang, Hongxin Wang
Cardiac fibrosis is considered an important pathological mechanism in the progression of cardiac remodeling and heart failure. Astragaloside IV (AsIV) is a major active ingredient in Astragalus membranaceus. In a preliminary experiment, it was demonstrated that this naturally occurring substance exhibited cardioprotective effects via preventing cardiomyocyte hypertrophy and apoptosis. The present study aimed to investigate the effects of AsIV on β‑adrenergic receptor (β‑AR)‑mediated cardiac fibrosis, and the associated mechanism...
April 2017: Molecular Medicine Reports
https://www.readbyqxmd.com/read/28235755/classical-and-novel-pharmacological-insights-offered-by-the-simple-chick-cardiomyocyte-cell-culture-model-a-valuable-teaching-aid-and-a-primer-for-real-research
#11
N S Freestone, C L S Sam
The chick embryo cardiomyocyte model of cell culture is a staple technique in many physiology and pharmacology laboratories. Despite the relative simplicity, robustness, and reproducibility inherent in this model, it can be used in a variety of ways to yield important new insights that help facilitate student understanding of underlying physiological and pharmacological concepts as well as, more generally, the scientific method. Using this model, this paper will show real data obtained by undergraduate students in the authors' laboratories...
March 1, 2017: Advances in Physiology Education
https://www.readbyqxmd.com/read/28231545/aqueous-extract-of-cortex-dictamni-protects-h9c2-cardiomyocytes-from-hypoxia-reoxygenation-induced-oxidative-stress-and-apoptosis-by-pi3k-akt-signaling-pathway
#12
Lin Li, Yunfeng Zhou, Yanlin Li, Lili Wang, Lan Sun, Lidong Zhou, Hiderori Arai, Yun Qi, Yang Xu
Ischemia-reperfusion injury is the major manifestation of ischemic heart disease, which facilitates cardiac arrhythmias, heart failure and death. Oxidative stress and apoptosis have been involved in the pathogenesis of myocardial ischemia-reperfusion injury. Modern pharmacological studies have indicated that the extracts and active compounds of Cortex Dictamni exhibit antioxidative and cardiovascular protective activities. This study was designed to investigate the protective effect of aqueous extract of Cortex Dictamni (CDAE) on regulating hypoxia/reoxygenation (H/R)-induced cardiomyocytes oxidative stress and apoptosis...
February 20, 2017: Biomedicine & Pharmacotherapy, Biomédecine & Pharmacothérapie
https://www.readbyqxmd.com/read/28230276/dissection-of-regulatory-elements-during-direct-conversion-of-somatic-cells-into-neurons
#13
Tahereh Soleimani, Nafiseh Falsafi, Hossein Fallahi
A revolutionary approach that involves direct conversion of somatic cells into almost any other types of cells showed promising results for regenerative medicine. Currently, producing valuable cell types including neurons, cardiomyocytes and hepatocytes through direct conversion of somatic cells appear to be a feasible option for regenerative medicine. The process involves inducing the cells by chemical cocktails or by expression of different types of transcription factors. In this concept, in vitro neurogenesis considered to be able to produce neuron cells to replace damaged neurons especially in Alzheimer and Parkinson disease...
February 23, 2017: Journal of Cellular Biochemistry
https://www.readbyqxmd.com/read/28229413/cardiac-progenitor-cell-extraction-from-human-auricles
#14
Paolo Di Nardo, Francesca Pagliari
For many years, myocardial tissue has been considered terminally differentiated and, thus, incapable of regenerating. Recent studies have shown, instead, that cardiomyocytes, at least in part, are slowly substituted by new cells originating by precursor cells mostly embedded into the heart apex and in the atria. We have shown that an elective region of progenitor cell embedding is represented by the auricles, non-contractile atria appendages that can be easily sampled without harming the patient. The protocol here reported describes how from auricles a population of multipotent, cardiogenic cells can be isolated, cultured, and differentiated...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28229406/manipulating-the-proliferative-potential-of-cardiomyocytes-by-gene-transfer
#15
Giulia Prosdocimo, Mauro Giacca
In contrast to prenatal life, cardiomyocyte proliferation in mammals is rapidly blunted after birth; as a consequence, clinically significant cardiac regeneration does not occur in adulthood. Thus, the modulation of cardiomyocyte proliferation by gene transfer offers an invaluable opportunity to both understand the mechanisms regulating renewal of these cells in the fetus and identify novel strategies for myocardial repair.In this Chapter, we report an exhaustive protocol to isolate, culture, and manipulate the properties of neonatal ventricular rat cardiomyocytes by small RNA transfection or transduction with viral vectors based on the adeno-associated virus, which exhibit exquisite tropism for these cells...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28224719/cardiomyocyte-differentiation-promotes-cell-survival-during-nicotinamide-phosphoribosyltransferase-inhibition-through-increased-maintenance-of-cellular-energy-stores
#16
Erin M Kropp, Katarzyna A Broniowska, Matthew Waas, Alyssa Nycz, John A Corbett, Rebekah L Gundry
To address concerns regarding the tumorigenic potential of undifferentiated human pluripotent stem cells (hPSC) that may remain after in vitro differentiation and ultimately limit the broad use of hPSC-derivatives for therapeutics, we recently described a method to selectively eliminate tumorigenic hPSC from their progeny by inhibiting nicotinamide phosphoribosyltransferase (NAMPT). Limited exposure to NAMPT inhibitors selectively removes hPSC from hPSC-derived cardiomyocytes (hPSC-CM) and spares a wide range of differentiated cell types; yet, it remains unclear when and how cells acquire resistance to NAMPT inhibition during differentiation...
February 22, 2017: Stem Cells Translational Medicine
https://www.readbyqxmd.com/read/28223521/nondestructive-nanostraw-intracellular-sampling-for-longitudinal-cell-monitoring
#17
Yuhong Cao, Martin Hjort, Haodong Chen, Fikri Birey, Sergio A Leal-Ortiz, Crystal M Han, Juan G Santiago, Sergiu P Paşca, Joseph C Wu, Nicholas A Melosh
Here, we report a method for time-resolved, longitudinal extraction and quantitative measurement of intracellular proteins and mRNA from a variety of cell types. Cytosolic contents were repeatedly sampled from the same cell or population of cells for more than 5 d through a cell-culture substrate, incorporating hollow 150-nm-diameter nanostraws (NS) within a defined sampling region. Once extracted, the cellular contents were analyzed with conventional methods, including fluorescence, enzymatic assays (ELISA), and quantitative real-time PCR...
March 7, 2017: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/28210753/puerarin-7-o-glucuronide-a-water-soluble-puerarin-metabolite-prevents-angiotensin-ii-induced-cardiomyocyte-hypertrophy-by-reducing-oxidative-stress
#18
Ning Hou, Bin Cai, Cai-Wen Ou, Zhen-Hui Zhang, Xia-Wen Liu, Mu Yuan, Gan-Jian Zhao, Shi-Ming Liu, Long-Gen Xiong, Jian-Dong Luo, Cheng-Feng Luo, Min-Sheng Chen
This study aimed to investigate the anti-oxidant and anti-hypertrophic effects of puerarin-7-O-glucuronide, a water-soluble puerarin metabolite. The anti-oxidant effects of puerarin-7-O-glucuronide were assessed by measurement of intracellular superoxide levels, total superoxide dismutase (SOD) activity, total anti-oxidant capacity, and glutathione (GSH)/glutathione disulfide (GSSG) ratio in cultured neonatal rat cardiomyocytes (NRCMs) stimulated with the xanthine oxidase (XO)/xanthine (X) system or angiotensin II...
February 16, 2017: Naunyn-Schmiedeberg's Archives of Pharmacology
https://www.readbyqxmd.com/read/28197936/salvianolic-acid-a-protects-neonatal-cardiomyocytes-against-hypoxia-reoxygenation-induced-injury-by-preserving-mitochondrial-function-and-activating-akt-gsk-3%C3%AE-signals
#19
Xue-Li Li, Ji-Ping Fan, Jian-Xun Liu, Li-Na Liang
OBJECTIVE: To investigate the effects of salvianolic acid A (SAA) on cardiomyocyte apoptosis and mitochondrial dysfunction in response to hypoxia/reoxygenation (H/R) injury and to determine whether the Akt signaling pathway might play a role. METHODS: An in vitro model of H/R injury was used to study outcomes on primary cultured neonatal rat cardiomyocytes. The cardiomyocytes were treated with 12.5, 25, 50 μg/mL SAA at the beginning of hypoxia and reoxygenation, respectively...
February 15, 2017: Chinese Journal of Integrative Medicine
https://www.readbyqxmd.com/read/28196901/novel-variant-in-the-ank2-membrane-binding-domain-is-associated-with-ankyrin-b-syndrome-and-structural-heart-disease-in-a-first-nations-population-with-a-high-rate-of-long-qt-syndrome
#20
Leigh Anne Swayne, Nathaniel P Murphy, Sirisha Asuri, Lena Chen, Xiaoxue Xu, Sarah McIntosh, Chao Wang, Peter J Lancione, Jason D Roberts, Charles Kerr, Shubhayan Sanatani, Elizabeth Sherwin, Crystal F Kline, Mingjie Zhang, Peter J Mohler, Laura T Arbour
BACKGROUND: Long QT syndrome confers susceptibility to ventricular arrhythmia, predisposing to syncope, seizures, and sudden death. While rare globally, long QT syndrome is ≈15× more common in First Nations of Northern British Columbia largely because of a known mutation in KCNQ1. However, 2 large multigenerational families were affected, but negative for the known mutation. METHODS AND RESULTS: Long QT syndrome panel testing was carried out in the index case of each family, and clinical information was collected...
January 2017: Circulation. Cardiovascular Genetics
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