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Cardiomyocyte Cell Culture

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https://www.readbyqxmd.com/read/29141446/myocardial-reparative-functions-of-exosomes-from-mesenchymal-stem-cells-are-enhanced-by-hypoxia-treatment-of-the-cells-via-transferring-microrna-210-in-an-nsmase2-dependent-way
#1
Jinyun Zhu, Kai Lu, Ning Zhang, Yun Zhao, Qunchao Ma, Jian Shen, Yinuo Lin, Pingping Xiang, Yaoliang Tang, Xinyang Hu, Jinghai Chen, Wei Zhu, Keith A Webster, Jian'an Wang, Hong Yu
Hypoxia treatment enhances paracrine effect of mesenchymal stem cells (MSCs). The aim of this study was to investigate whether exosomes from hypoxia-treated MSCs (Exo(H)) are superior to those from normoxia-treated MSCs (Exo(N)) for myocardial repair. Mouse bone marrow-derived MSCs were cultured under hypoxia or normoxia for 24 h, and exosomes from conditioned media were intramyocardially injected into infarcted heart of C57BL/6 mouse. Exo(H) resulted in significantly higher survival, smaller scar size and better cardiac functions recovery...
November 16, 2017: Artificial Cells, Nanomedicine, and Biotechnology
https://www.readbyqxmd.com/read/29140569/single-cell-functional-analysis-of-stem-cell-derived-cardiomyocytes-on-micropatterned-flexible-substrates
#2
Jan David Kijlstra, Dongjian Hu, Peter van der Meer, Ibrahim J Domian
Human pluripotent stem-cell derived cardiomyocytes (hPSC-CMs) hold great promise for applications in human disease modeling, drug discovery, cardiotoxicity screening, and, ultimately, regenerative medicine. The ability to study multiple parameters of hPSC-CM function, such as contractile and electrical activity, calcium cycling, and force generation, is therefore of paramount importance. hPSC-CMs cultured on stiff substrates like glass or polystyrene do not have the ability to shorten during contraction, making them less suitable for the study of hPSC-CM contractile function...
November 15, 2017: Current Protocols in Stem Cell Biology
https://www.readbyqxmd.com/read/29133848/community-effect-of-cardiomyocytes-in-beating-rhythms-is-determined-by-stable-cells
#3
Tatsuya Hayashi, Tetsuji Tokihiro, Hiroki Kurihara, Kenji Yasuda
The community effect of cardiomyocytes was investigated in silico by the change in number and features of cells, as well as configurations of networks. The theoretical model was based on experimental data and accurately reproduced recently published experimental results regarding coupled cultured cardiomyocytes. We showed that the synchronised beating of two coupled cells was tuned not to the cell with a faster beating rate, but to the cell with a more stable rhythm. In a network of cardiomyocytes, a cell with low fluctuation, but not a hight frequency, became a pacemaker and stabilised the beating rhythm...
November 13, 2017: Scientific Reports
https://www.readbyqxmd.com/read/29133820/early-postnatal-cardiomyocyte-proliferation-requires-high-oxidative-energy-metabolism
#4
Ana Elisa Teófilo Saturi de Carvalho, Vinícius Bassaneze, Maria Fernanda Forni, Aline Alfonso Keusseyan, Alicia Juliana Kowaltowski, José Eduardo Krieger
Cardiac energy metabolism must cope with early postnatal changes in tissue oxygen tensions, hemodynamics, and cell proliferation to sustain development. Here, we tested the hypothesis that proliferating neonatal cardiomyocytes are dependent on high oxidative energy metabolism. We show that energy-related gene expression does not correlate with functional oxidative measurements in the developing heart. Gene expression analysis suggests a gradual overall upregulation of oxidative-related genes and pathways, whereas functional assessment in both cardiac tissue and cultured cardiomyocytes indicated that oxidative metabolism decreases between the first and seventh days after birth...
November 13, 2017: Scientific Reports
https://www.readbyqxmd.com/read/29130959/ginsenoside-rg1-protects-cardiomyocytes-against-hypoxia-reoxygenation-injury-via-activation-of-nrf2-ho-1-signaling-and-inhibition-of-jnk
#5
Qianhui Li, Yin Xiang, Yu Chen, Yong Tang, Yachen Zhang
BACKGROUND/AIMS: Excessive reactive oxygen species (ROS) disturb the physiology of H9c2 cells, which is regarded as a major cause of H9c2 cardiomyocyte apoptosis. Ginsenoside Rg1 is the main active extract of ginseng, which has important antioxidant properties in various cell models. This project investigated the role of ginsenoside Rg1 in hypoxia/reoxygenation (H/R)-induced oxidative stress injury in cultured H9c2 cells to reveal the underlying signaling pathways. METHODS: H9c2 cells were pretreated with ginsenoside Rg1 for 12 h before exposure to H/R...
November 3, 2017: Cellular Physiology and Biochemistry
https://www.readbyqxmd.com/read/29130958/propofol-through-upregulating-caveolin-3-attenuates-post-hypoxic-mitochondrial-damage-and-cell-death-in-h9c2-cardiomyocytes-during-hyperglycemia
#6
Fan Deng, Shuang Wang, Liangqing Zhang, Xiang Xie, Shuyun Cai, Haobo Li, Gui-Ling Xie, Hui-Lai Miao, Changmin Yang, Xin Liu, Zhengyuan Xia
BACKGROUND/AIMS: Hearts from diabetic subjects are susceptible to myocardial ischemia reperfusion (I/R) injury. Propofol has been shown to protect against myocardial I/R injury due to its antioxidant properties while the underlying mechanism remained incompletely understood. Thus, this study aimed to determine whether or not propofol could attenuate myocardial I/R injury by attenuating mitochondrial dysfunction/damage through upregulating Caveolin (Cav)-3 under hyperglycemia. METHODS: Cultured rat cardiomyocyte H9C2 cells were subjected to hypoxia/reoxygenation (H/R) in the absence or presence of propofol under high glucose (HG), and cell viability, lactate dehydrogenase (LDH) and mitochondrial viability as well as creatine kinase-MB (CK-MB), cardiac troponin I (cTnI) and intracellular adenosine triphosphate (ATP) content were measured with colorimetric Enzyme-Linked Immunosorbent Assays...
November 9, 2017: Cellular Physiology and Biochemistry
https://www.readbyqxmd.com/read/29128638/ulk1-regulated-autophagy-a-mechanism-in-cellular-protection-for-aldh2-against-hyperglycemia
#7
Min Liu, Songhe Lu, Wei He, Le Zhang, Ying Ma, Ping Lv, Meijuan Ma, Wenjun Yu, Jiaxing Wang, Mingming Zhang, Yingmei Zhang, Yan Li
Mitochondrial aldehyde dehydrogenase 2 (ALDH2), an important enzyme in the elimination of toxic aldehydes, is involved in cardioprotection against diabetes mellitus. This study was designed to examine the mechanism behind ALDH2-offered protection against high glucose exposure with a focus on autophagy. H9C2 cells were cultured with normal or high glucose medium in the presence or absence of the ALDH2 agonist Alda-1. GFP-LC3 puncta and immunofluorescence were employed to assess autophagosome formation. Western blotting was applied to evaluate autophagy protein markers Atg5, LC3, p62, ULK1 phosphorylation and ALDH2...
November 8, 2017: Toxicology Letters
https://www.readbyqxmd.com/read/29127009/nicorandil-alleviates-myocardial-injury-and-post-infarction-cardiac-remodeling-by-inhibiting-mst1
#8
Shanjie Wang, Yanhong Fan, Xinyu Feng, Chuang Sun, Zhaofeng Shi, Tian Li, Jianjun Lv, Zhi Yang, Zhijing Zhao, Dongdong Sun
BACKGROUND: Cardiomyocyte autophagy and apoptosis are crucial events underlying the development of cardiac abnormalities and dysfunction after myocardial infarction (MI). A better understanding of the cell signaling pathways involved in cardiac remodeling may support the development of new therapeutic strategies for the treatment of heart failure (HF) after MI. METHODS: A cardiac MI injury model was constructed by ligating the left anterior descending (LAD) coronary artery...
November 7, 2017: Biochemical and Biophysical Research Communications
https://www.readbyqxmd.com/read/29126879/mechanical-stretch-increases-l-type-calcium-channel-stability-in-cardiomyocytes-through-a-polycystin-1-akt-dependent-mechanism
#9
A Córdova-Casanova, I Olmedo, J A Riquelme, G Barrientos, G Sánchez, T G Gillette, S Lavandero, M Chiong, P Donoso, Z Pedrozo
The L-type calcium channel (LTCC) is an important determinant of cardiac contractility. Therefore, changes in LTCC activity or protein levels could be expected to affect cardiac function. Several studies describing LTCC regulation are available, but only a few examine LTCC protein stability. Polycystin-1 (PC1) is a mechanosensor that regulates heart contractility and is involved in mechanical stretch-induced cardiac hypertrophy. PC1 was originally described as an unconventional Gi/o protein-coupled receptor in renal cells...
November 7, 2017: Biochimica et Biophysica Acta
https://www.readbyqxmd.com/read/29125993/substrate-and-mechanotransduction-influence-serca2a-localization-in-human-pluripotent-stem-cell-derived-cardiomyocytes-affecting-functional-performance
#10
Sebastian Martewicz, Elena Serena, Susi Zatti, Gordon Keller, Nicola Elvassore
Physical cues are major determinants of cellular phenotype and evoke physiological and pathological responses on cell structure and function. Cellular models aim to recapitulate basic functional features of their in vivo counterparts or tissues in order to be of use in in vitro disease modeling or drug screening and testing. Understanding how culture systems affect in vitro development of human pluripotent stem cell (hPSC)-derivatives allows optimization of cellular human models and gives insight in the processes involved in their structural organization and function...
October 16, 2017: Stem Cell Research
https://www.readbyqxmd.com/read/29116112/a-comparison-of-mrna-sequencing-with-random-primed-and-3-directed-libraries
#11
Yuguang Xiong, Magali Soumillon, Jie Wu, Jens Hansen, Bin Hu, Johan G C van Hasselt, Gomathi Jayaraman, Ryan Lim, Mehdi Bouhaddou, Loren Ornelas, Jim Bochicchio, Lindsay Lenaeus, Jennifer Stocksdale, Jaehee Shim, Emilda Gomez, Dhruv Sareen, Clive Svendsen, Leslie M Thompson, Milind Mahajan, Ravi Iyengar, Eric A Sobie, Evren U Azeloglu, Marc R Birtwistle
Creating a cDNA library for deep mRNA sequencing (mRNAseq) is generally done by random priming, creating multiple sequencing fragments along each transcript. A 3'-end-focused library approach cannot detect differential splicing, but has potentially higher throughput at a lower cost, along with the ability to improve quantification by using transcript molecule counting with unique molecular identifiers (UMI) that correct PCR bias. Here, we compare an implementation of such a 3'-digital gene expression (3'-DGE) approach with "conventional" random primed mRNAseq...
November 7, 2017: Scientific Reports
https://www.readbyqxmd.com/read/29115435/human-umbilical-cord-mesenchymal-stem-cells-alleviate-interstitial-fibrosis-and-cardiac-dysfunction-in-a-dilated-cardiomyopathy-rat-model-by-inhibiting-tnf%C3%A2-%C3%AE-and-tgf%C3%A2-%C3%AE-1-erk1-2-signaling-pathways
#12
Changyi Zhang, Guichi Zhou, Yezeng Chen, Sizheng Liu, Fen Chen, Lichun Xie, Wei Wang, Yonggang Zhang, Tianyou Wang, Xiulan Lai, Lian Ma
Dilated cardiomyopathy (DCM) is a disease of the heart characterized by pathological remodeling, including patchy interstitial fibrosis and degeneration of cardiomyocytes. In the present study, the beneficial role of human umbilical cord‑derived mesenchymal stem cells (HuMSCs) derived from Wharton's jelly was evaluated in the myosin‑induced rat model of DCM. Male Lewis rats (aged 8‑weeks) were injected with porcine myosin to induce DCM. Cultured HuMSCs (1x106 cells/rat) were intravenously injected 28 days after myosin injection and the effects on myocardial fibrosis and the underlying signaling pathways were investigated and compared with vehicle‑injected and negative control rats...
October 26, 2017: Molecular Medicine Reports
https://www.readbyqxmd.com/read/29115402/ampk-activation-serves-a-critical-role-in-mitochondria-quality-control-via-modulating-mitophagy-in-the-heart-under-chronic-hypoxia
#13
Huagang Zhang, Bo Liu, Tianbo Li, Yun Zhu, Guiping Luo, Yunhan Jiang, Fuqin Tang, Zhao Jian, Yingbin Xiao
Mitochondrial biogenesis is one of the generally accepted regulatory mechanisms in the heart under chronic hypoxia. The precise quantity and quality control of mitochondria is critical for the survival and function of cardiomyocytes. Mitochondrial autophagy, also known as mitophagy, which selectively eliminates dysfunctional and unwanted mitochondria, is the most important type of mitochondrial quality control. However, the detailed molecular mechanisms of mitophagy in cardiomyocytes have been largely undefined...
October 26, 2017: International Journal of Molecular Medicine
https://www.readbyqxmd.com/read/29113299/microrna-210-suppresses-glucocorticoid-receptor-expression-in-response-to-hypoxia-in-fetal-rat-cardiomyocytes
#14
Shannalee R Martinez, Qingyi Ma, Chiranjib Dasgupta, Xianmei Meng, Lubo Zhang
Hypoxia is a common intrauterine stressor, often resulting in intrauterine growth restriction and increased risk for cardiovascular disease later in life. The aim of this work was to test the hypothesis that microRNA-210 (miR-210) mediates the detrimental suppression of glucocorticoid receptor (GR) in response to hypoxia in fetal rat cardiomyocytes. Cardiomyocytes isolated from gestational day 21 Sprague Dawley fetal rats showed increased miR-210 levels and reduced GR abundance after exposure to ex vivo hypoxia (1% O2)...
October 6, 2017: Oncotarget
https://www.readbyqxmd.com/read/29108785/semaphorin7a-aggravates-coxsackievirusb3-induced-viral-myocarditis-by-increasing-%C3%AE-1%C3%AE-1-integrin-macrophages-and-subsequent-enhanced-inflammatory-response
#15
Xuejie Wu, Yawen Meng, Chao Wang, Yan Yue, Chunsheng Dong, Sidong Xiong
Semaphorin7A (Sema7A) has been reported to play various roles in nerve axon growth, tumor suppression, and tissue remodeling, as well as regulation of intestinal inflammation diseases. Viral myocarditis (VMC) characterized by viral-myocardial-cell necrosis and inflammatory cell infiltration is a common clinical disease of the cardiovascular system. However, the role of Sema7A in coxsackievirus B3 (CVB3)-induced VMC has not been reported. In this study, we generated an acute VMC mouse model by CVB3 infection, and manipulated Sema7A expression by in vivo polyethyleneimine-mediated Sema7A down-regulation...
November 3, 2017: Journal of Molecular and Cellular Cardiology
https://www.readbyqxmd.com/read/29100414/the-ros-nf-%C3%AE%C2%BAb-nr4a2-pathway-is-involved-in-h2o2-induced-apoptosis-of-resident-cardiac-stem-cells-via-autophagy
#16
Xingxing Shi, Wenjing Li, Honghong Liu, Deling Yin, Jing Zhao
Cardiac stem cells (CSCs)-based therapy provides a promising avenue for the management of ischemic heart diseases. However, engrafted CSCs are subjected to acute cell apoptosis in the ischemic microenvironment. Here, stem cell antigen 1 positive (Sca-1(+)) CSCs proved to own therapy potential were cultured and treated with H2O2 to mimic the ischemia situation. As autophagy inhibitor, 3-methyladenine (3MA), inhibited H2O2-induced CSCs apoptosis, thus we demonstrated that H2O2 induced autophagy-dependent apoptosis in CSCs, and continued to find key proteins responsible for the crosstalk between autophagy and apoptosis...
September 29, 2017: Oncotarget
https://www.readbyqxmd.com/read/29096746/overexpression-of-microrna-133b-reduces-myocardial-injuries-in-children-with-viral-myocarditis-by-targeting-rab27b-gene
#17
Y Zhang, L Sun, H Sun, X Liu, X Luo, C Li, D Sun, T Li
The present study is to measure the expression of microRNA (miRNA or miR)-133b in circulating blood of children with viral myocarditis before and after drug treatment, and to investigate its relationship with the severity of myocardial lesions. A total of 36 children patients with viral myocarditis who received treatments at our hospital between June 2014 and June 2016 were enrolled in the present study, including 21 boys and 15 girls (age range, 9 months - 16 years).Quantitative real-time polymerase chain reaction was used to determine the expression of miR-133b in peripheral blood of patients and cardiomyocytes infected with CVB3...
October 31, 2017: Cellular and Molecular Biology
https://www.readbyqxmd.com/read/29093370/-development-of-targeted-pharmacotherapy-for-cardiovascular-disease
#18
Yasufumi Katanasaka
 Heart and cardiovascular diseases are the leading causes of death in the world. Heart failure (HF) in particular is becoming a serious widespread medical issue, especially following various stresses such as myocardial infarction and hemodynamic overload. One pathological cardiac change in HF is left ventricular hypertrophy (LVH). LVH is associated with increased risk for HF; however, no drug therapy for LVH has yet been developed. During the development of LVH, gene expression is altered in cardiomyocytes through transcription factors, co-activators, and histone modifications...
2017: Yakugaku Zasshi: Journal of the Pharmaceutical Society of Japan
https://www.readbyqxmd.com/read/29091577/laminin-laminin-entactin-and-extracellular-matrix-are-equally-appropriate-adhesive-substrates-for-isolated-adult-rat-cardiomyocyte-culture-and-experimentation
#19
D Lumkwana, A Botha, E Samodien, S Hanser, J Lopes
Although techniques for isolating and culturing adult cardiomyocytes were developed four decades ago, it still remains a challenge to isolate high yields of healthy viable cardiomyocytes, to maintain them in culture, and to use them successfully in experiments. This is due to the difficulty in deciding which adhesive substrate and buffer composition to use. Therefore this study aimed to (1) identify a robust experimental buffer to sustain survival of cultured adult rat cardiomyocytes (ARCMs) during control normoxic conditions, and (2) to identify an adhesive substrate that provides optimal cell adherence, not only during normoxia, but especially during simulated ischemia-reperfusion (SIR) experiments...
November 1, 2017: Cell Adhesion & Migration
https://www.readbyqxmd.com/read/29078393/hspb7-is-indispensable-for-heart-development-by-modulating-actin-filament-assembly
#20
Tongbin Wu, Yongxin Mu, Julius Bogomolovas, Xi Fang, Jennifer Veevers, Roberta B Nowak, Christopher T Pappas, Carol C Gregorio, Sylvia M Evans, Velia M Fowler, Ju Chen
Small heat shock protein HSPB7 is highly expressed in the heart. Several mutations within HSPB7 are associated with dilated cardiomyopathy and heart failure in human patients. However, the precise role of HSPB7 in the heart is still unclear. In this study, we generated global as well as cardiac-specific HSPB7 KO mouse models and found that loss of HSPB7 globally or specifically in cardiomyocytes resulted in embryonic lethality before embryonic day 12.5. Using biochemical and cell culture assays, we identified HSPB7 as an actin filament length regulator that repressed actin polymerization by binding to monomeric actin...
November 7, 2017: Proceedings of the National Academy of Sciences of the United States of America
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