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  • 92
    ATCC at 1 mouse atrial cardiomyocyte tumor lineage
    At 1 Mouse Atrial Cardiomyocyte Tumor Lineage, supplied by ATCC, used in various techniques. Bioz Stars score: 92/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore at 1 mouse atrial cardiomyocyte tumour lineage
    At 1 Mouse Atrial Cardiomyocyte Tumour Lineage, supplied by Millipore, used in various techniques. Bioz Stars score: 97/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Thermo Fisher neonatal rat mouse cardiomyocyte isolation kit
    Molecular events of ROS-dependent PKCδ activation involved in the AGE-BSA-induced <t>cardiomyocyte</t> apoptosis. PKCδ activation is involved in the regulation of AGE-BSA-induced cell apoptosis via ROS production and may play a key role in the development of cardiac mitochondrial dysfunction in rats with diabetes or obesity.
    Neonatal Rat Mouse Cardiomyocyte Isolation Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 22 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    The Jackson Laboratory cardiomyocyte specific mercremer mice
    New cardiomyocytes are derived from pre-existing cardiomyocytes during aging a) Experimental strategy. <t>MerCreMer</t> + /ZEG + mice (n=4) treated for 2 wks with 4OH-tamoxifen to induce <t>cardiomyocyte-specific</t> GFP expression. 15 N-thymidine administered continuously during 10 wk chase, then cycling cells identified by 15 N-labeling. New cardiomyocytes ( 15 N + ) derived from preexisting cardiomyocytes should express GFP at a rate similar to surrounding quiescent ( 15 N − ) cardiomyocytes. New cardiomyocytes ( 15 N + ) derived from progenitors should be GFP − . b) Left: 15 N: 14 N HSI image demonstrating 15 N-thymidine labeled cardiomyocyte nucleus (white asterisk) and 15 N-labeled non-cardiomyocyte (white arrow). Right: Immunofluorescent image showing 15 N-labeled cardiomyocyte is GFP + . Scale bar = 15μm.
    Cardiomyocyte Specific Mercremer Mice, supplied by The Jackson Laboratory, used in various techniques. Bioz Stars score: 85/100, based on 27 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Miltenyi Biotec neonatal cardiomyocyte isolation kit mouse
    ( a , b ) (left) The long-term culture of hIPSC-derived cardiac myocytes yields cell assemblies similar to the culture of mouse primary cardiac myocytes, giving rise to the assumption that such stem cell populations can be used to substitute for primary animal-derived specimens; ( c , d ) Different from the well-ordered  α -actinin apparatus, the connexin-43 distribution does not recapitulate the cell interface accumulation as seen in neonatal cardiac myocyte culture; ( e ) Functional (left) and defect (right) cell ensembles imaged by SEM. The obvious finding that the well-spread cell ensemble is a functional one while the defect one releases surface contacts and reaches a bulk appearance does not absolutely correlate with a loss of contractility. A loss of surface-mediated organization can be regarded inside the bulk assembly. Consequently, spatially-random contracting bulks can be observed; ( f , b ) (right) Aging effects of the contractile apparatus after long-term cultivation. The regular  α -actinin organization with a ladder appearance with pronounced orientation along cells’ longitudes is substituted by a random orientation; ( g ) Ensembles of cardiac myocytes beat synchronously, indicating electrical excitation coupling. While the direction of contraction is random on a plain surface (right), the line pattern-induced cell ensemble organization results in temporally synchronized and spatially-organized contractions. This fulfills a fundamental prerequisite of cardiac tissue function.
    Neonatal Cardiomyocyte Isolation Kit Mouse, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 98/100, based on 18 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore monoclonal anti alpha actinin sarcomeric antibody
    ( a , b ) (left) The long-term culture of hIPSC-derived cardiac myocytes yields cell assemblies similar to the culture of mouse primary cardiac myocytes, giving rise to the assumption that such stem cell populations can be used to substitute for primary animal-derived specimens; ( c , d ) Different from the well-ordered  α -actinin apparatus, the connexin-43 distribution does not recapitulate the cell interface accumulation as seen in neonatal cardiac myocyte culture; ( e ) Functional (left) and defect (right) cell ensembles imaged by SEM. The obvious finding that the well-spread cell ensemble is a functional one while the defect one releases surface contacts and reaches a bulk appearance does not absolutely correlate with a loss of contractility. A loss of surface-mediated organization can be regarded inside the bulk assembly. Consequently, spatially-random contracting bulks can be observed; ( f , b ) (right) Aging effects of the contractile apparatus after long-term cultivation. The regular  α -actinin organization with a ladder appearance with pronounced orientation along cells’ longitudes is substituted by a random orientation; ( g ) Ensembles of cardiac myocytes beat synchronously, indicating electrical excitation coupling. While the direction of contraction is random on a plain surface (right), the line pattern-induced cell ensemble organization results in temporally synchronized and spatially-organized contractions. This fulfills a fundamental prerequisite of cardiac tissue function.
    Monoclonal Anti Alpha Actinin Sarcomeric Antibody, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 2806 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ScienCell c57bl 6 mouse ventricular cardiomyoctes
    ( a , b ) (left) The long-term culture of hIPSC-derived cardiac myocytes yields cell assemblies similar to the culture of mouse primary cardiac myocytes, giving rise to the assumption that such stem cell populations can be used to substitute for primary animal-derived specimens; ( c , d ) Different from the well-ordered  α -actinin apparatus, the connexin-43 distribution does not recapitulate the cell interface accumulation as seen in neonatal cardiac myocyte culture; ( e ) Functional (left) and defect (right) cell ensembles imaged by SEM. The obvious finding that the well-spread cell ensemble is a functional one while the defect one releases surface contacts and reaches a bulk appearance does not absolutely correlate with a loss of contractility. A loss of surface-mediated organization can be regarded inside the bulk assembly. Consequently, spatially-random contracting bulks can be observed; ( f , b ) (right) Aging effects of the contractile apparatus after long-term cultivation. The regular  α -actinin organization with a ladder appearance with pronounced orientation along cells’ longitudes is substituted by a random orientation; ( g ) Ensembles of cardiac myocytes beat synchronously, indicating electrical excitation coupling. While the direction of contraction is random on a plain surface (right), the line pattern-induced cell ensemble organization results in temporally synchronized and spatially-organized contractions. This fulfills a fundamental prerequisite of cardiac tissue function.
    C57bl 6 Mouse Ventricular Cardiomyoctes, supplied by ScienCell, used in various techniques. Bioz Stars score: 92/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc adult mouse cardiomyocytes
    ( a , b ) (left) The long-term culture of hIPSC-derived cardiac myocytes yields cell assemblies similar to the culture of mouse primary cardiac myocytes, giving rise to the assumption that such stem cell populations can be used to substitute for primary animal-derived specimens; ( c , d ) Different from the well-ordered  α -actinin apparatus, the connexin-43 distribution does not recapitulate the cell interface accumulation as seen in neonatal cardiac myocyte culture; ( e ) Functional (left) and defect (right) cell ensembles imaged by SEM. The obvious finding that the well-spread cell ensemble is a functional one while the defect one releases surface contacts and reaches a bulk appearance does not absolutely correlate with a loss of contractility. A loss of surface-mediated organization can be regarded inside the bulk assembly. Consequently, spatially-random contracting bulks can be observed; ( f , b ) (right) Aging effects of the contractile apparatus after long-term cultivation. The regular  α -actinin organization with a ladder appearance with pronounced orientation along cells’ longitudes is substituted by a random orientation; ( g ) Ensembles of cardiac myocytes beat synchronously, indicating electrical excitation coupling. While the direction of contraction is random on a plain surface (right), the line pattern-induced cell ensemble organization results in temporally synchronized and spatially-organized contractions. This fulfills a fundamental prerequisite of cardiac tissue function.
    Adult Mouse Cardiomyocytes, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 92/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    IonOptix adult mouse cardiomyocytes
    ( a , b ) (left) The long-term culture of hIPSC-derived cardiac myocytes yields cell assemblies similar to the culture of mouse primary cardiac myocytes, giving rise to the assumption that such stem cell populations can be used to substitute for primary animal-derived specimens; ( c , d ) Different from the well-ordered  α -actinin apparatus, the connexin-43 distribution does not recapitulate the cell interface accumulation as seen in neonatal cardiac myocyte culture; ( e ) Functional (left) and defect (right) cell ensembles imaged by SEM. The obvious finding that the well-spread cell ensemble is a functional one while the defect one releases surface contacts and reaches a bulk appearance does not absolutely correlate with a loss of contractility. A loss of surface-mediated organization can be regarded inside the bulk assembly. Consequently, spatially-random contracting bulks can be observed; ( f , b ) (right) Aging effects of the contractile apparatus after long-term cultivation. The regular  α -actinin organization with a ladder appearance with pronounced orientation along cells’ longitudes is substituted by a random orientation; ( g ) Ensembles of cardiac myocytes beat synchronously, indicating electrical excitation coupling. While the direction of contraction is random on a plain surface (right), the line pattern-induced cell ensemble organization results in temporally synchronized and spatially-organized contractions. This fulfills a fundamental prerequisite of cardiac tissue function.
    Adult Mouse Cardiomyocytes, supplied by IonOptix, used in various techniques. Bioz Stars score: 88/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cardiac Dimensions cardiomyocyte mr knockout mice
    Important GPCR-related molecular pathways involved in crosstalk with the MR in cardiac myocytes. Aldo: Aldosterone; P: Phosphorylation. Solid arrows indicate direct effect, whereas dotted arrows indicate indirect (through additional intermediate proteins) effect. See text for details and for all other molecular acronym descriptions.
    Cardiomyocyte Mr Knockout Mice, supplied by Cardiac Dimensions, used in various techniques. Bioz Stars score: 92/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Merck & Co hl1 immortalised mouse cardiomyocyte cell line
    Important GPCR-related molecular pathways involved in crosstalk with the MR in cardiac myocytes. Aldo: Aldosterone; P: Phosphorylation. Solid arrows indicate direct effect, whereas dotted arrows indicate indirect (through additional intermediate proteins) effect. See text for details and for all other molecular acronym descriptions.
    Hl1 Immortalised Mouse Cardiomyocyte Cell Line, supplied by Merck & Co, used in various techniques. Bioz Stars score: 93/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    97
    Millipore monoclonal anti actin alpha sarcomeric antibody
    Important GPCR-related molecular pathways involved in crosstalk with the MR in cardiac myocytes. Aldo: Aldosterone; P: Phosphorylation. Solid arrows indicate direct effect, whereas dotted arrows indicate indirect (through additional intermediate proteins) effect. See text for details and for all other molecular acronym descriptions.
    Monoclonal Anti Actin Alpha Sarcomeric Antibody, supplied by Millipore, used in various techniques. Bioz Stars score: 97/100, based on 550 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc adult mouse 538 cardiomyocytes
    Important GPCR-related molecular pathways involved in crosstalk with the MR in cardiac myocytes. Aldo: Aldosterone; P: Phosphorylation. Solid arrows indicate direct effect, whereas dotted arrows indicate indirect (through additional intermediate proteins) effect. See text for details and for all other molecular acronym descriptions.
    Adult Mouse 538 Cardiomyocytes, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 84/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Charles River Laboratories adult mouse primary cardiomyocytes adult c57bl 6 mice
    Important GPCR-related molecular pathways involved in crosstalk with the MR in cardiac myocytes. Aldo: Aldosterone; P: Phosphorylation. Solid arrows indicate direct effect, whereas dotted arrows indicate indirect (through additional intermediate proteins) effect. See text for details and for all other molecular acronym descriptions.
    Adult Mouse Primary Cardiomyocytes Adult C57bl 6 Mice, supplied by Charles River Laboratories, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher adult rat mouse cardiomyocyte isolation kit
    Bone marrow cells adopt <t>cardiomyocyte</t> fates within the heart. ( A ) Flow cytometry analysis demonstrating the DsRed+/troponin+ population. ( B ) Percentage of troponin+ cells in MI injured and sham surgery hearts. ( C ) Percentage of bone marrow-derived troponin+ cells in MI injured and sham surgery hearts. ( D ) Percentage of troponin+ cells due to fusion (GFP+/DsRed+) or de novo formation (GFP−/DsRed+). ( E ) Bone marrow-derived troponin+ cells outside of infarct area that co-express DsRed and troponin. Scale bar, 25 μm. ( F ) Sorted cardiomyocytes. The superior cell expresses troponin, DsRed and GFP and likely arose from cell fusion. The inferior cell only expresses troponin and DsRed and likely arose from de novo generation. Scale bar, 50 μm.
    Adult Rat Mouse Cardiomyocyte Isolation Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 92/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Warner Instruments hypoxia reoxygenation model mouse cardiomyocytes
    Effect of CypD deletion on hypercontracture, mPTP opening and cell death after 45 min hypoxia followed by <t>reoxygenation</t> in isolated adult mouse <t>cardiomyocytes.</t> Cardiomyocytes were isolated from wild type (WT) and CypD knock-out (KO) mice, were co-loaded with calcein, CoCl 2 and propidium iodide (PI) and were subjected to 45 min hypoxia and then reoxygenated. Images were collected every min during the first 5 min of reoxygenation and then every 5 min. ( a ) Percentage of cells exhibiting hypercontracture at the end of hypoxia and at different reoxygenation times. ( b ) Time course of mPTP opening and cell death during reoxygenation. Fluorescence was normalised to 100% of the maximal values. Traces from 3 to 5 experiments were averaged.
    Hypoxia Reoxygenation Model Mouse Cardiomyocytes, supplied by Warner Instruments, used in various techniques. Bioz Stars score: 91/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher human cardiomyocyte immunocytochemistry kit
    Effect of CypD deletion on hypercontracture, mPTP opening and cell death after 45 min hypoxia followed by <t>reoxygenation</t> in isolated adult mouse <t>cardiomyocytes.</t> Cardiomyocytes were isolated from wild type (WT) and CypD knock-out (KO) mice, were co-loaded with calcein, CoCl 2 and propidium iodide (PI) and were subjected to 45 min hypoxia and then reoxygenated. Images were collected every min during the first 5 min of reoxygenation and then every 5 min. ( a ) Percentage of cells exhibiting hypercontracture at the end of hypoxia and at different reoxygenation times. ( b ) Time course of mPTP opening and cell death during reoxygenation. Fluorescence was normalised to 100% of the maximal values. Traces from 3 to 5 experiments were averaged.
    Human Cardiomyocyte Immunocytochemistry Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 27 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher troponin t cardiac isoform ab 1 mouse monoclonal antibody
    Effect of CypD deletion on hypercontracture, mPTP opening and cell death after 45 min hypoxia followed by <t>reoxygenation</t> in isolated adult mouse <t>cardiomyocytes.</t> Cardiomyocytes were isolated from wild type (WT) and CypD knock-out (KO) mice, were co-loaded with calcein, CoCl 2 and propidium iodide (PI) and were subjected to 45 min hypoxia and then reoxygenated. Images were collected every min during the first 5 min of reoxygenation and then every 5 min. ( a ) Percentage of cells exhibiting hypercontracture at the end of hypoxia and at different reoxygenation times. ( b ) Time course of mPTP opening and cell death during reoxygenation. Fluorescence was normalised to 100% of the maximal values. Traces from 3 to 5 experiments were averaged.
    Troponin T Cardiac Isoform Ab 1 Mouse Monoclonal Antibody, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 633 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    84
    Cosmo Bio cardiomyocyte culture mouse ventricular cardiomyocytes
    Effect of CypD deletion on hypercontracture, mPTP opening and cell death after 45 min hypoxia followed by <t>reoxygenation</t> in isolated adult mouse <t>cardiomyocytes.</t> Cardiomyocytes were isolated from wild type (WT) and CypD knock-out (KO) mice, were co-loaded with calcein, CoCl 2 and propidium iodide (PI) and were subjected to 45 min hypoxia and then reoxygenated. Images were collected every min during the first 5 min of reoxygenation and then every 5 min. ( a ) Percentage of cells exhibiting hypercontracture at the end of hypoxia and at different reoxygenation times. ( b ) Time course of mPTP opening and cell death during reoxygenation. Fluorescence was normalised to 100% of the maximal values. Traces from 3 to 5 experiments were averaged.
    Cardiomyocyte Culture Mouse Ventricular Cardiomyocytes, supplied by Cosmo Bio, used in various techniques. Bioz Stars score: 84/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ATCC cardiomyocytes mouse c2c12 myoblasts
    GLP-1 and GLP-1(9-36) increased endosomal and sarcolemmal Glut4 in hyperlipidemic <t>cardiomyocytes.</t> Glut4 expression in endosome- (e) and sarcolemmal (s)-enriched fractions of a HG- and b HF-stimulated <t>C2C12.</t> Some cells were pre-treated with GLP-1 or GLP-1(9-36). *p
    Cardiomyocytes Mouse C2c12 Myoblasts, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher cardiomyocyte specific mouse monoclonal anti α actinin
    GLP-1 and GLP-1(9-36) increased endosomal and sarcolemmal Glut4 in hyperlipidemic <t>cardiomyocytes.</t> Glut4 expression in endosome- (e) and sarcolemmal (s)-enriched fractions of a HG- and b HF-stimulated <t>C2C12.</t> Some cells were pre-treated with GLP-1 or GLP-1(9-36). *p
    Cardiomyocyte Specific Mouse Monoclonal Anti α Actinin, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    The Jackson Laboratory cardiomyocyte specific myh6 mercremer mice
    GLP-1 and GLP-1(9-36) increased endosomal and sarcolemmal Glut4 in hyperlipidemic <t>cardiomyocytes.</t> Glut4 expression in endosome- (e) and sarcolemmal (s)-enriched fractions of a HG- and b HF-stimulated <t>C2C12.</t> Some cells were pre-treated with GLP-1 or GLP-1(9-36). *p
    Cardiomyocyte Specific Myh6 Mercremer Mice, supplied by The Jackson Laboratory, used in various techniques. Bioz Stars score: 93/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Takeda cardiomyocyte specific stat3 conditional knockout mice stat3 flox mice
    <t>STAT3</t> was activated in <t>cardiomyocytes</t> at EAM3w. ( a ) Heart homogenates at EAM0w and EAM3w were subjected to immunoblotting with anti-phosphorylated STAT3 (pSTAT3) (Y705), anti-pSTAT3 (S727), anti-pAkt, anti-pERK and anti-GAPDH antibodies. Blots were reprobed with anti-total STAT3 (tSTAT3), anti-tAkt and anti-tERK antibodies. The full-length blots are presented in Supplementary Figure S3 . ( b – e ) The band intensity was measured with ImageJ and normalized to that of GAPDH. Data are shown as fold increase relative to 0w. n = 4 mice for 0w; 6 mice for 3w. ( f ) Heart sections were immunostained with anti-pSTAT3 (Y705) and anti-cTnI antibodies at the indicated time points after EAM induction. Left: representative images of pSTAT3 (Y705) + cTnI + cells at EAM3w are shown. Arrows: pSTAT3 (Y705) + nuclei in cTnI + cells. Scale bar: 50 μm. Right: pSTAT3 (Y705) + cTnI + cells in the inflamed region were counted and shown as percentage in cTnI + cells. n = 3 mice for each group. ( g ) The expression of IL-6, IL-11, LIF, OSM, CLCF1 and CT-1 transcripts was quantified at EAM0w and EAM3w by quantitative RT-PCR. The expression of these genes was normalized to that of gapdh and shown as fold increase relative to 0w. n = 5 mice for 0w; 8 mice for 3w and 5w. ( b – e ) Welch’s t -test; ( f ) Kruskal-Wallis test; ( g ) Steel-Dwass test. ** P
    Cardiomyocyte Specific Stat3 Conditional Knockout Mice Stat3 Flox Mice, supplied by Takeda, used in various techniques. Bioz Stars score: 91/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Molecular events of ROS-dependent PKCδ activation involved in the AGE-BSA-induced cardiomyocyte apoptosis. PKCδ activation is involved in the regulation of AGE-BSA-induced cell apoptosis via ROS production and may play a key role in the development of cardiac mitochondrial dysfunction in rats with diabetes or obesity.

    Journal: Aging and Disease

    Article Title: Pkcδ Activation is Involved in ROS-Mediated Mitochondrial Dysfunction and Apoptosis in Cardiomyocytes Exposed to Advanced Glycation End Products (Ages)

    doi: 10.14336/AD.2017.0924

    Figure Lengend Snippet: Molecular events of ROS-dependent PKCδ activation involved in the AGE-BSA-induced cardiomyocyte apoptosis. PKCδ activation is involved in the regulation of AGE-BSA-induced cell apoptosis via ROS production and may play a key role in the development of cardiac mitochondrial dysfunction in rats with diabetes or obesity.

    Article Snippet: Neonatal rat ventricular myocytes primary culture NRVMs were prepared and cultured using a Neonatal Rat/Mouse Cardiomyocyte Isolation Kit (nc-6031; Cellutron Life Technology, Baltimore, MD, USA) which was previously described [ ].

    Techniques: Activation Assay

    AGE-BSA-induced cardiomyocyte apoptosis is mediated through ROS-dependent PKCδ activation NRVM and H9c2 cells were exposed to AGE-BSA (300 μg/ml) for 24 h. Cells were co-treated with bryostatin 1, a PKCδ activator (100 nM), rottlerin, a PKCδ inhibitor (3 μM), NAC (500 μM), Rote (0.1 μM), Apo (10 μM) or siPKCδ (1 μg). ( A ) Expression and phosphorylation of PKCδ and ( B, C, D ) apoptosis-related proteins were examined by western blot analyses. These are cropped blots, full-length blots of PKCδ and pPKCδ are presented in Suppl. Figure S5 . β-Actin was used as a loading control. N-acetylcysteine, NAC; Rote, rotenone; APO, apocynin; SC, scramble.

    Journal: Aging and Disease

    Article Title: Pkcδ Activation is Involved in ROS-Mediated Mitochondrial Dysfunction and Apoptosis in Cardiomyocytes Exposed to Advanced Glycation End Products (Ages)

    doi: 10.14336/AD.2017.0924

    Figure Lengend Snippet: AGE-BSA-induced cardiomyocyte apoptosis is mediated through ROS-dependent PKCδ activation NRVM and H9c2 cells were exposed to AGE-BSA (300 μg/ml) for 24 h. Cells were co-treated with bryostatin 1, a PKCδ activator (100 nM), rottlerin, a PKCδ inhibitor (3 μM), NAC (500 μM), Rote (0.1 μM), Apo (10 μM) or siPKCδ (1 μg). ( A ) Expression and phosphorylation of PKCδ and ( B, C, D ) apoptosis-related proteins were examined by western blot analyses. These are cropped blots, full-length blots of PKCδ and pPKCδ are presented in Suppl. Figure S5 . β-Actin was used as a loading control. N-acetylcysteine, NAC; Rote, rotenone; APO, apocynin; SC, scramble.

    Article Snippet: Neonatal rat ventricular myocytes primary culture NRVMs were prepared and cultured using a Neonatal Rat/Mouse Cardiomyocyte Isolation Kit (nc-6031; Cellutron Life Technology, Baltimore, MD, USA) which was previously described [ ].

    Techniques: Activation Assay, Expressing, Western Blot

    AGE-BSA-induced cardiomyocyte apoptosis is mediated through PKCδ activation. ( A ) The diagram depicts the domain organization of GFP-PKCδ. The GFP-PKCδ derivatives, including the wild-type (WT) and the kinase-deficient mutant (KD; K376R). Cells were treated with AGE-BSA (300 μg/ml) and ( B ) rottlerin (1-5 μM) or ( C ) PKCδ silencing. ( D ) Cells were transfected with GFP-fused PKCδ (GFP PKCδ-WT) at different doses as indicated and with ( E ) 1 μg rottlerin (1-5 μM). ( F G ) H9c2 cells or ( H ) neonatal rat ventricular myocytes (NRVM) were exposed to AGEs (300 μg/ml) with or without (GFP PKCδ-KD) transfection or transfected with GFP PKCδ-WT in the presence of rottlerin (3 μM) or not. These are cropped blots, full-length blots of PKCδ and pPKCδ are presented in Suppl. Figure S4 . SC, scramble; WT, wild type; KD, kinase-deficient; All the proteins were analyzed by western blotting using β-actin as a loading control.

    Journal: Aging and Disease

    Article Title: Pkcδ Activation is Involved in ROS-Mediated Mitochondrial Dysfunction and Apoptosis in Cardiomyocytes Exposed to Advanced Glycation End Products (Ages)

    doi: 10.14336/AD.2017.0924

    Figure Lengend Snippet: AGE-BSA-induced cardiomyocyte apoptosis is mediated through PKCδ activation. ( A ) The diagram depicts the domain organization of GFP-PKCδ. The GFP-PKCδ derivatives, including the wild-type (WT) and the kinase-deficient mutant (KD; K376R). Cells were treated with AGE-BSA (300 μg/ml) and ( B ) rottlerin (1-5 μM) or ( C ) PKCδ silencing. ( D ) Cells were transfected with GFP-fused PKCδ (GFP PKCδ-WT) at different doses as indicated and with ( E ) 1 μg rottlerin (1-5 μM). ( F G ) H9c2 cells or ( H ) neonatal rat ventricular myocytes (NRVM) were exposed to AGEs (300 μg/ml) with or without (GFP PKCδ-KD) transfection or transfected with GFP PKCδ-WT in the presence of rottlerin (3 μM) or not. These are cropped blots, full-length blots of PKCδ and pPKCδ are presented in Suppl. Figure S4 . SC, scramble; WT, wild type; KD, kinase-deficient; All the proteins were analyzed by western blotting using β-actin as a loading control.

    Article Snippet: Neonatal rat ventricular myocytes primary culture NRVMs were prepared and cultured using a Neonatal Rat/Mouse Cardiomyocyte Isolation Kit (nc-6031; Cellutron Life Technology, Baltimore, MD, USA) which was previously described [ ].

    Techniques: Activation Assay, Mutagenesis, Transfection, Western Blot

    New cardiomyocytes are derived from pre-existing cardiomyocytes during aging a) Experimental strategy. MerCreMer + /ZEG + mice (n=4) treated for 2 wks with 4OH-tamoxifen to induce cardiomyocyte-specific GFP expression. 15 N-thymidine administered continuously during 10 wk chase, then cycling cells identified by 15 N-labeling. New cardiomyocytes ( 15 N + ) derived from preexisting cardiomyocytes should express GFP at a rate similar to surrounding quiescent ( 15 N − ) cardiomyocytes. New cardiomyocytes ( 15 N + ) derived from progenitors should be GFP − . b) Left: 15 N: 14 N HSI image demonstrating 15 N-thymidine labeled cardiomyocyte nucleus (white asterisk) and 15 N-labeled non-cardiomyocyte (white arrow). Right: Immunofluorescent image showing 15 N-labeled cardiomyocyte is GFP + . Scale bar = 15μm.

    Journal: Nature

    Article Title: Mammalian Heart Renewal by Preexisting Cardiomyocytes

    doi: 10.1038/nature11682

    Figure Lengend Snippet: New cardiomyocytes are derived from pre-existing cardiomyocytes during aging a) Experimental strategy. MerCreMer + /ZEG + mice (n=4) treated for 2 wks with 4OH-tamoxifen to induce cardiomyocyte-specific GFP expression. 15 N-thymidine administered continuously during 10 wk chase, then cycling cells identified by 15 N-labeling. New cardiomyocytes ( 15 N + ) derived from preexisting cardiomyocytes should express GFP at a rate similar to surrounding quiescent ( 15 N − ) cardiomyocytes. New cardiomyocytes ( 15 N + ) derived from progenitors should be GFP − . b) Left: 15 N: 14 N HSI image demonstrating 15 N-thymidine labeled cardiomyocyte nucleus (white asterisk) and 15 N-labeled non-cardiomyocyte (white arrow). Right: Immunofluorescent image showing 15 N-labeled cardiomyocyte is GFP + . Scale bar = 15μm.

    Article Snippet: We generated double transgenic Mer-CreMer-ZEG male mice by crossbreeding cardiomyocyte-specific MerCreMer mice and ZEG mice (Jackson Laboratory). β-galactosidase-GFP is under the control of a cytomegalovirus (CMV) enhancer/chicken actin promoter (Actb ); the background strain was C57BL/6J (N7).

    Techniques: Derivative Assay, Mouse Assay, Expressing, Labeling

    Myocardial injury stimulates division of pre-existing cardiomyocytes a) Myocardial infarction (MI) leads to extensive DNA synthesis within and adjacent to scar (arrows). MerCreMer + /ZEG + mice were treated for 2 wks with 4OH-tamoxifen to induce cardiomyocyte-specific GFP expression before MI or sham surgery, then 15 N-thymidine administered continuously for 8wks. Mosaics of 70 60×60μm MIMS tiles. Trichrome stained adjacent section (far right) shows scar. Scale bars = 90μm. b) 15 N-thymidine labeled cardiomyocyte nucleus (white arrows) from MI border region. Immunofluorescent staining demonstrates that the cardiomyocyte is GFP + . Scale bars = 10 μm. c) Mean % 15 N + cardiomyocyte nuclei after MI (n=4) in the scar border region compared to sham operated mice (n=3). Mean% ± S.E.M.

    Journal: Nature

    Article Title: Mammalian Heart Renewal by Preexisting Cardiomyocytes

    doi: 10.1038/nature11682

    Figure Lengend Snippet: Myocardial injury stimulates division of pre-existing cardiomyocytes a) Myocardial infarction (MI) leads to extensive DNA synthesis within and adjacent to scar (arrows). MerCreMer + /ZEG + mice were treated for 2 wks with 4OH-tamoxifen to induce cardiomyocyte-specific GFP expression before MI or sham surgery, then 15 N-thymidine administered continuously for 8wks. Mosaics of 70 60×60μm MIMS tiles. Trichrome stained adjacent section (far right) shows scar. Scale bars = 90μm. b) 15 N-thymidine labeled cardiomyocyte nucleus (white arrows) from MI border region. Immunofluorescent staining demonstrates that the cardiomyocyte is GFP + . Scale bars = 10 μm. c) Mean % 15 N + cardiomyocyte nuclei after MI (n=4) in the scar border region compared to sham operated mice (n=3). Mean% ± S.E.M.

    Article Snippet: We generated double transgenic Mer-CreMer-ZEG male mice by crossbreeding cardiomyocyte-specific MerCreMer mice and ZEG mice (Jackson Laboratory). β-galactosidase-GFP is under the control of a cytomegalovirus (CMV) enhancer/chicken actin promoter (Actb ); the background strain was C57BL/6J (N7).

    Techniques: DNA Synthesis, Mouse Assay, Expressing, Staining, Labeling

    ( a , b ) (left) The long-term culture of hIPSC-derived cardiac myocytes yields cell assemblies similar to the culture of mouse primary cardiac myocytes, giving rise to the assumption that such stem cell populations can be used to substitute for primary animal-derived specimens; ( c , d ) Different from the well-ordered  α -actinin apparatus, the connexin-43 distribution does not recapitulate the cell interface accumulation as seen in neonatal cardiac myocyte culture; ( e ) Functional (left) and defect (right) cell ensembles imaged by SEM. The obvious finding that the well-spread cell ensemble is a functional one while the defect one releases surface contacts and reaches a bulk appearance does not absolutely correlate with a loss of contractility. A loss of surface-mediated organization can be regarded inside the bulk assembly. Consequently, spatially-random contracting bulks can be observed; ( f , b ) (right) Aging effects of the contractile apparatus after long-term cultivation. The regular  α -actinin organization with a ladder appearance with pronounced orientation along cells’ longitudes is substituted by a random orientation; ( g ) Ensembles of cardiac myocytes beat synchronously, indicating electrical excitation coupling. While the direction of contraction is random on a plain surface (right), the line pattern-induced cell ensemble organization results in temporally synchronized and spatially-organized contractions. This fulfills a fundamental prerequisite of cardiac tissue function.

    Journal: Journal of Functional Biomaterials

    Article Title: Tissue-Mimicking Geometrical Constraints Stimulate Tissue-Like Constitution and Activity of Mouse Neonatal and Human-Induced Pluripotent Stem Cell-Derived Cardiac Myocytes

    doi: 10.3390/jfb7010001

    Figure Lengend Snippet: ( a , b ) (left) The long-term culture of hIPSC-derived cardiac myocytes yields cell assemblies similar to the culture of mouse primary cardiac myocytes, giving rise to the assumption that such stem cell populations can be used to substitute for primary animal-derived specimens; ( c , d ) Different from the well-ordered α -actinin apparatus, the connexin-43 distribution does not recapitulate the cell interface accumulation as seen in neonatal cardiac myocyte culture; ( e ) Functional (left) and defect (right) cell ensembles imaged by SEM. The obvious finding that the well-spread cell ensemble is a functional one while the defect one releases surface contacts and reaches a bulk appearance does not absolutely correlate with a loss of contractility. A loss of surface-mediated organization can be regarded inside the bulk assembly. Consequently, spatially-random contracting bulks can be observed; ( f , b ) (right) Aging effects of the contractile apparatus after long-term cultivation. The regular α -actinin organization with a ladder appearance with pronounced orientation along cells’ longitudes is substituted by a random orientation; ( g ) Ensembles of cardiac myocytes beat synchronously, indicating electrical excitation coupling. While the direction of contraction is random on a plain surface (right), the line pattern-induced cell ensemble organization results in temporally synchronized and spatially-organized contractions. This fulfills a fundamental prerequisite of cardiac tissue function.

    Article Snippet: Cell Culture For isolation of primary neonatal mouse cardiac myocytes, the Neonatal Heart Dissociation Kit and the Neonatal Cardiomyocyte Isolation Kit for mice (No. 130-098-373, No. 130-100-825, Miltenyi Biotec, Bergisch Gladbach, Germany) are used according to the manufacturer recommendations.

    Techniques: Derivative Assay, Functional Assay

    Fluorescence ( a ) and bright-field ( b ) light microscopy recordings of a typical murine cardiomyocyte specimen situation. A multicellular ensemble in near-confluent cell density is orientated along the polymer line pattern series; ( a , c ) (red and green inserts) Dual color overlay of the intracellular α -actinin (red) distribution and the intercellular connexin-43 distribution (green). The red signal closely follows the cell shapes, indicating that all cells are densely filled with α -actinin. Image (c) represents a higher magnification of the green rectangle with the α -actinin recording; ( d , e ) (blue insert) Distribution of the gap junction protein connexin-43. The pearl chain-like accumulations are located at the cell interfaces inside the cell contacts. A more homogeneous distribution of small signals all over the cytosol indicates non-recruited protein; ( f , g ) SEM recording of murine cardiac myocytes cultivated on line-patterned substrate. Vital cells are orientated along the fillets, while a randomly-distributed residual of rounded cells indicates the loss of cells during preparation from heart tissue; ( f ) Near confluent inoculation does not impede the organizing influence of line patterns, but it reduces the ratio of long cell protrusions. Such protrusions are more apparent at low cell densities (g). The golden overlay in (g) hints at the chamfer component of the surface pattern. An abstract sketch of the alternating chamfer to line pattern is given in Figure 1 .

    Journal: Journal of Functional Biomaterials

    Article Title: Tissue-Mimicking Geometrical Constraints Stimulate Tissue-Like Constitution and Activity of Mouse Neonatal and Human-Induced Pluripotent Stem Cell-Derived Cardiac Myocytes

    doi: 10.3390/jfb7010001

    Figure Lengend Snippet: Fluorescence ( a ) and bright-field ( b ) light microscopy recordings of a typical murine cardiomyocyte specimen situation. A multicellular ensemble in near-confluent cell density is orientated along the polymer line pattern series; ( a , c ) (red and green inserts) Dual color overlay of the intracellular α -actinin (red) distribution and the intercellular connexin-43 distribution (green). The red signal closely follows the cell shapes, indicating that all cells are densely filled with α -actinin. Image (c) represents a higher magnification of the green rectangle with the α -actinin recording; ( d , e ) (blue insert) Distribution of the gap junction protein connexin-43. The pearl chain-like accumulations are located at the cell interfaces inside the cell contacts. A more homogeneous distribution of small signals all over the cytosol indicates non-recruited protein; ( f , g ) SEM recording of murine cardiac myocytes cultivated on line-patterned substrate. Vital cells are orientated along the fillets, while a randomly-distributed residual of rounded cells indicates the loss of cells during preparation from heart tissue; ( f ) Near confluent inoculation does not impede the organizing influence of line patterns, but it reduces the ratio of long cell protrusions. Such protrusions are more apparent at low cell densities (g). The golden overlay in (g) hints at the chamfer component of the surface pattern. An abstract sketch of the alternating chamfer to line pattern is given in Figure 1 .

    Article Snippet: Cell Culture For isolation of primary neonatal mouse cardiac myocytes, the Neonatal Heart Dissociation Kit and the Neonatal Cardiomyocyte Isolation Kit for mice (No. 130-098-373, No. 130-100-825, Miltenyi Biotec, Bergisch Gladbach, Germany) are used according to the manufacturer recommendations.

    Techniques: Fluorescence, Light Microscopy

    ( a ) Survey of a group of neonatal murine cardiac myocytes on a smooth glass substrate. The cells are stained for nuclei (blue), gap junctions via connexin-43 immunostaining (green) and the contractile apparatus via  α -actinin immuno-staining (red); ( b ) (insert in (a))Exclusive presentation of the  α -actinin signal component; ( c ) Exclusive presentation of the connexin-43 signal component. Besides the accumulation along pearl chain-like structures located along the intercellular borders, a faint non-specific signal is also visible inside the nuclei due to a cross-over in the acquisition channel. ( d )–( f ) are SEM recordings of neonatal cardiac myocytes cultivated on plain glass surface. The cells are flat and polygonal with an irregular polygonal shape.

    Journal: Journal of Functional Biomaterials

    Article Title: Tissue-Mimicking Geometrical Constraints Stimulate Tissue-Like Constitution and Activity of Mouse Neonatal and Human-Induced Pluripotent Stem Cell-Derived Cardiac Myocytes

    doi: 10.3390/jfb7010001

    Figure Lengend Snippet: ( a ) Survey of a group of neonatal murine cardiac myocytes on a smooth glass substrate. The cells are stained for nuclei (blue), gap junctions via connexin-43 immunostaining (green) and the contractile apparatus via α -actinin immuno-staining (red); ( b ) (insert in (a))Exclusive presentation of the α -actinin signal component; ( c ) Exclusive presentation of the connexin-43 signal component. Besides the accumulation along pearl chain-like structures located along the intercellular borders, a faint non-specific signal is also visible inside the nuclei due to a cross-over in the acquisition channel. ( d )–( f ) are SEM recordings of neonatal cardiac myocytes cultivated on plain glass surface. The cells are flat and polygonal with an irregular polygonal shape.

    Article Snippet: Cell Culture For isolation of primary neonatal mouse cardiac myocytes, the Neonatal Heart Dissociation Kit and the Neonatal Cardiomyocyte Isolation Kit for mice (No. 130-098-373, No. 130-100-825, Miltenyi Biotec, Bergisch Gladbach, Germany) are used according to the manufacturer recommendations.

    Techniques: Staining, Immunostaining

    Important GPCR-related molecular pathways involved in crosstalk with the MR in cardiac myocytes. Aldo: Aldosterone; P: Phosphorylation. Solid arrows indicate direct effect, whereas dotted arrows indicate indirect (through additional intermediate proteins) effect. See text for details and for all other molecular acronym descriptions.

    Journal: International Journal of Molecular Sciences

    Article Title: Novel Insights into the Crosstalk between Mineralocorticoid Receptor and G Protein-Coupled Receptors in Heart Adverse Remodeling and Disease

    doi: 10.3390/ijms19123764

    Figure Lengend Snippet: Important GPCR-related molecular pathways involved in crosstalk with the MR in cardiac myocytes. Aldo: Aldosterone; P: Phosphorylation. Solid arrows indicate direct effect, whereas dotted arrows indicate indirect (through additional intermediate proteins) effect. See text for details and for all other molecular acronym descriptions.

    Article Snippet: Cardiomyocyte MR-knockout mice have normal systolic and diastolic functions and cardiac dimensions [ ].

    Techniques:

    Bone marrow cells adopt cardiomyocyte fates within the heart. ( A ) Flow cytometry analysis demonstrating the DsRed+/troponin+ population. ( B ) Percentage of troponin+ cells in MI injured and sham surgery hearts. ( C ) Percentage of bone marrow-derived troponin+ cells in MI injured and sham surgery hearts. ( D ) Percentage of troponin+ cells due to fusion (GFP+/DsRed+) or de novo formation (GFP−/DsRed+). ( E ) Bone marrow-derived troponin+ cells outside of infarct area that co-express DsRed and troponin. Scale bar, 25 μm. ( F ) Sorted cardiomyocytes. The superior cell expresses troponin, DsRed and GFP and likely arose from cell fusion. The inferior cell only expresses troponin and DsRed and likely arose from de novo generation. Scale bar, 50 μm.

    Journal: Journal of cardiovascular translational research

    Article Title: SDF 1-alpha attenuates myocardial injury without altering the direct contribution of circulating cells

    doi: 10.1007/s12265-017-9772-y

    Figure Lengend Snippet: Bone marrow cells adopt cardiomyocyte fates within the heart. ( A ) Flow cytometry analysis demonstrating the DsRed+/troponin+ population. ( B ) Percentage of troponin+ cells in MI injured and sham surgery hearts. ( C ) Percentage of bone marrow-derived troponin+ cells in MI injured and sham surgery hearts. ( D ) Percentage of troponin+ cells due to fusion (GFP+/DsRed+) or de novo formation (GFP−/DsRed+). ( E ) Bone marrow-derived troponin+ cells outside of infarct area that co-express DsRed and troponin. Scale bar, 25 μm. ( F ) Sorted cardiomyocytes. The superior cell expresses troponin, DsRed and GFP and likely arose from cell fusion. The inferior cell only expresses troponin and DsRed and likely arose from de novo generation. Scale bar, 50 μm.

    Article Snippet: The Adult Rat/Mouse Cardiomyocyte Isolation Kit (Cat# AC-7031 from Cellutron Life Technologies, Baltimore, MD, USA) was used to digest the heart.

    Techniques: Flow Cytometry, Cytometry, Derivative Assay

    Effect of CypD deletion on hypercontracture, mPTP opening and cell death after 45 min hypoxia followed by reoxygenation in isolated adult mouse cardiomyocytes. Cardiomyocytes were isolated from wild type (WT) and CypD knock-out (KO) mice, were co-loaded with calcein, CoCl 2 and propidium iodide (PI) and were subjected to 45 min hypoxia and then reoxygenated. Images were collected every min during the first 5 min of reoxygenation and then every 5 min. ( a ) Percentage of cells exhibiting hypercontracture at the end of hypoxia and at different reoxygenation times. ( b ) Time course of mPTP opening and cell death during reoxygenation. Fluorescence was normalised to 100% of the maximal values. Traces from 3 to 5 experiments were averaged.

    Journal: Scientific Reports

    Article Title: Ca2+ ionophores are not suitable for inducing mPTP opening in murine isolated adult cardiac myocytes

    doi: 10.1038/s41598-017-04618-4

    Figure Lengend Snippet: Effect of CypD deletion on hypercontracture, mPTP opening and cell death after 45 min hypoxia followed by reoxygenation in isolated adult mouse cardiomyocytes. Cardiomyocytes were isolated from wild type (WT) and CypD knock-out (KO) mice, were co-loaded with calcein, CoCl 2 and propidium iodide (PI) and were subjected to 45 min hypoxia and then reoxygenated. Images were collected every min during the first 5 min of reoxygenation and then every 5 min. ( a ) Percentage of cells exhibiting hypercontracture at the end of hypoxia and at different reoxygenation times. ( b ) Time course of mPTP opening and cell death during reoxygenation. Fluorescence was normalised to 100% of the maximal values. Traces from 3 to 5 experiments were averaged.

    Article Snippet: Hypoxia-reoxygenation model Mouse cardiomyocytes were placed into a thermostated (37 °C) chamber (Warner Instruments Inc, Connecticut) which was mounted on the stage of an IX81 Olympus microscope (Olympus, Rungis, France) and were perfused with the Tyrode’s solution at a rate of 0.5 ml/min.

    Techniques: Isolation, Knock-Out, Mouse Assay, Fluorescence

    GLP-1 and GLP-1(9-36) increased endosomal and sarcolemmal Glut4 in hyperlipidemic cardiomyocytes. Glut4 expression in endosome- (e) and sarcolemmal (s)-enriched fractions of a HG- and b HF-stimulated C2C12. Some cells were pre-treated with GLP-1 or GLP-1(9-36). *p

    Journal: Cardiovascular Diabetology

    Article Title: Sitagliptin improved glucose assimilation in detriment of fatty-acid utilization in experimental type-II diabetes: role of GLP-1 isoforms in Glut4 receptor trafficking

    doi: 10.1186/s12933-017-0643-2

    Figure Lengend Snippet: GLP-1 and GLP-1(9-36) increased endosomal and sarcolemmal Glut4 in hyperlipidemic cardiomyocytes. Glut4 expression in endosome- (e) and sarcolemmal (s)-enriched fractions of a HG- and b HF-stimulated C2C12. Some cells were pre-treated with GLP-1 or GLP-1(9-36). *p

    Article Snippet: Cultured cardiomyocytes Mouse C2C12 myoblasts (ATCC, USA) were kindly given by Dr. Konhilas (University of Arizona, USA), and maintained in DMEM supplemented with 9% foetal calf serum, 5 mM d -glucose, 50 U/ml penicillin, and 50 μg/ml streptomycin.

    Techniques: Expressing

    STAT3 was activated in cardiomyocytes at EAM3w. ( a ) Heart homogenates at EAM0w and EAM3w were subjected to immunoblotting with anti-phosphorylated STAT3 (pSTAT3) (Y705), anti-pSTAT3 (S727), anti-pAkt, anti-pERK and anti-GAPDH antibodies. Blots were reprobed with anti-total STAT3 (tSTAT3), anti-tAkt and anti-tERK antibodies. The full-length blots are presented in Supplementary Figure S3 . ( b – e ) The band intensity was measured with ImageJ and normalized to that of GAPDH. Data are shown as fold increase relative to 0w. n = 4 mice for 0w; 6 mice for 3w. ( f ) Heart sections were immunostained with anti-pSTAT3 (Y705) and anti-cTnI antibodies at the indicated time points after EAM induction. Left: representative images of pSTAT3 (Y705) + cTnI + cells at EAM3w are shown. Arrows: pSTAT3 (Y705) + nuclei in cTnI + cells. Scale bar: 50 μm. Right: pSTAT3 (Y705) + cTnI + cells in the inflamed region were counted and shown as percentage in cTnI + cells. n = 3 mice for each group. ( g ) The expression of IL-6, IL-11, LIF, OSM, CLCF1 and CT-1 transcripts was quantified at EAM0w and EAM3w by quantitative RT-PCR. The expression of these genes was normalized to that of gapdh and shown as fold increase relative to 0w. n = 5 mice for 0w; 8 mice for 3w and 5w. ( b – e ) Welch’s t -test; ( f ) Kruskal-Wallis test; ( g ) Steel-Dwass test. ** P

    Journal: Scientific Reports

    Article Title: Adult murine cardiomyocytes exhibit regenerative activity with cell cycle reentry through STAT3 in the healing process of myocarditis

    doi: 10.1038/s41598-017-01426-8

    Figure Lengend Snippet: STAT3 was activated in cardiomyocytes at EAM3w. ( a ) Heart homogenates at EAM0w and EAM3w were subjected to immunoblotting with anti-phosphorylated STAT3 (pSTAT3) (Y705), anti-pSTAT3 (S727), anti-pAkt, anti-pERK and anti-GAPDH antibodies. Blots were reprobed with anti-total STAT3 (tSTAT3), anti-tAkt and anti-tERK antibodies. The full-length blots are presented in Supplementary Figure S3 . ( b – e ) The band intensity was measured with ImageJ and normalized to that of GAPDH. Data are shown as fold increase relative to 0w. n = 4 mice for 0w; 6 mice for 3w. ( f ) Heart sections were immunostained with anti-pSTAT3 (Y705) and anti-cTnI antibodies at the indicated time points after EAM induction. Left: representative images of pSTAT3 (Y705) + cTnI + cells at EAM3w are shown. Arrows: pSTAT3 (Y705) + nuclei in cTnI + cells. Scale bar: 50 μm. Right: pSTAT3 (Y705) + cTnI + cells in the inflamed region were counted and shown as percentage in cTnI + cells. n = 3 mice for each group. ( g ) The expression of IL-6, IL-11, LIF, OSM, CLCF1 and CT-1 transcripts was quantified at EAM0w and EAM3w by quantitative RT-PCR. The expression of these genes was normalized to that of gapdh and shown as fold increase relative to 0w. n = 5 mice for 0w; 8 mice for 3w and 5w. ( b – e ) Welch’s t -test; ( f ) Kruskal-Wallis test; ( g ) Steel-Dwass test. ** P

    Article Snippet: Cardiomyocyte-specific STAT3 conditional knockout mice STAT3 flox mice were a generous gift from Dr. Kiyoshi Takeda, Graduate School of Medicine, Osaka University.

    Techniques: Mouse Assay, Expressing, Quantitative RT-PCR

    STAT3 gene ablation suppressed the frequency of proliferative cardiomyocytes with impaired myocardial restoration from EAM. ( a ) Tamoxifen was injected to double transgenic mice with α -MHC-MerCreMer and STAT3 flox to ablate STAT3 gene in cardiomyocytes before EAM induction. ( b ) Cardiomyocytes isolated from STAT3fl/fl and STAT3cKO hearts at EAM3w were subjected to immunoblotting with anti-pSTAT3 (Y705), anti-tSTAT3 and anti-GAPDH antibodies. The full-length blots are presented in Supplementary Figure S5 . ( c ) The band intensity was measured with ImageJ and normalized to that of GAPDH. Data are shown as fold increase relative to fl/fl. n = 7 mice for fl/fl; 4 mice for cKO. ( d ) Representative STAT3fl/fl and STAT3cKO hearts at EAM5w. Scale bar: 2 mm. ( e ) The ratio of heart weight to body weight was calculated for the indicated groups. n = 5 mice for fl/fl 0w and cKO 0w; 7 mice for fl/fl 5w; 14 mice for cKO 5w. ( f ) Fractional shortening of STAT3fl/fl and STAT3cKO mice was evaluated by echocardiography at EAM0w and EAM5w. n = 9 mice for each group. ( g ) HE staining was performed for heart sections from STAT3fl/fl and STAT3cKO mice at EAM5w. Left: representative images are shown. Scale bar: 100 μm. Right: injured area was measured. Data were from 7 mice for each group. ( h ) The number of cardiomyocytes in post-inflamed areas of STAT3fl/fl and STAT3cKO hearts was counted at EAM5w and the density was calculated. More than 18,000 myocytes were counted from 7 mice for each group. ( i ) Heart sections from STAT3fl/fl and STAT3cKO mice at EAM3w were immunostained for Ki-67 and MHC. Ki-67 + MHC + cells in the inflamed region were counted and shown as percentage in MHC + cells. n = 5 mice for fl/fl; 8 mice for cKO. ( j ) Heart sections from STAT3fl/fl and STAT3cKO mice at EAM3w were immunostained for Aurora B and cTnI. Aurora B + cTnI + cells in the inflamed region were counted and shown as percentage in cTnI + cells. n = 5 mice for fl/fl; 8 mice for cKO. ( k ) BrdU was intraperitoneally injected four times into STAT3fl/fl and STAT3cKO mice at EAM3w. Heart sections were immunostained for BrdU and MHC 24 hours after the last injection. BrdU + MHC + cells in the inflamed region were counted and shown as percentage in MHC + cells. n = 5 mice for fl/fl; 8 mice for cKO. ( l ) Cardiomyocytes were isolated from STAT3fl/fl or STAT3cKO mice at EAM5w and stained with anti-α-actinin antibody and DAPI. A total of 1296 cardiomyocytes from 5 STAT3fl/fl hearts and 1367 cardiomyocytes from 5 STAT3cKO hearts were classified according to the number of nuclei. ( c , g – l ) Welch’s t -test; ( e ) two-way factorial ANOVA; ( f ) two-way repeated measures ANOVA. * P

    Journal: Scientific Reports

    Article Title: Adult murine cardiomyocytes exhibit regenerative activity with cell cycle reentry through STAT3 in the healing process of myocarditis

    doi: 10.1038/s41598-017-01426-8

    Figure Lengend Snippet: STAT3 gene ablation suppressed the frequency of proliferative cardiomyocytes with impaired myocardial restoration from EAM. ( a ) Tamoxifen was injected to double transgenic mice with α -MHC-MerCreMer and STAT3 flox to ablate STAT3 gene in cardiomyocytes before EAM induction. ( b ) Cardiomyocytes isolated from STAT3fl/fl and STAT3cKO hearts at EAM3w were subjected to immunoblotting with anti-pSTAT3 (Y705), anti-tSTAT3 and anti-GAPDH antibodies. The full-length blots are presented in Supplementary Figure S5 . ( c ) The band intensity was measured with ImageJ and normalized to that of GAPDH. Data are shown as fold increase relative to fl/fl. n = 7 mice for fl/fl; 4 mice for cKO. ( d ) Representative STAT3fl/fl and STAT3cKO hearts at EAM5w. Scale bar: 2 mm. ( e ) The ratio of heart weight to body weight was calculated for the indicated groups. n = 5 mice for fl/fl 0w and cKO 0w; 7 mice for fl/fl 5w; 14 mice for cKO 5w. ( f ) Fractional shortening of STAT3fl/fl and STAT3cKO mice was evaluated by echocardiography at EAM0w and EAM5w. n = 9 mice for each group. ( g ) HE staining was performed for heart sections from STAT3fl/fl and STAT3cKO mice at EAM5w. Left: representative images are shown. Scale bar: 100 μm. Right: injured area was measured. Data were from 7 mice for each group. ( h ) The number of cardiomyocytes in post-inflamed areas of STAT3fl/fl and STAT3cKO hearts was counted at EAM5w and the density was calculated. More than 18,000 myocytes were counted from 7 mice for each group. ( i ) Heart sections from STAT3fl/fl and STAT3cKO mice at EAM3w were immunostained for Ki-67 and MHC. Ki-67 + MHC + cells in the inflamed region were counted and shown as percentage in MHC + cells. n = 5 mice for fl/fl; 8 mice for cKO. ( j ) Heart sections from STAT3fl/fl and STAT3cKO mice at EAM3w were immunostained for Aurora B and cTnI. Aurora B + cTnI + cells in the inflamed region were counted and shown as percentage in cTnI + cells. n = 5 mice for fl/fl; 8 mice for cKO. ( k ) BrdU was intraperitoneally injected four times into STAT3fl/fl and STAT3cKO mice at EAM3w. Heart sections were immunostained for BrdU and MHC 24 hours after the last injection. BrdU + MHC + cells in the inflamed region were counted and shown as percentage in MHC + cells. n = 5 mice for fl/fl; 8 mice for cKO. ( l ) Cardiomyocytes were isolated from STAT3fl/fl or STAT3cKO mice at EAM5w and stained with anti-α-actinin antibody and DAPI. A total of 1296 cardiomyocytes from 5 STAT3fl/fl hearts and 1367 cardiomyocytes from 5 STAT3cKO hearts were classified according to the number of nuclei. ( c , g – l ) Welch’s t -test; ( e ) two-way factorial ANOVA; ( f ) two-way repeated measures ANOVA. * P

    Article Snippet: Cardiomyocyte-specific STAT3 conditional knockout mice STAT3 flox mice were a generous gift from Dr. Kiyoshi Takeda, Graduate School of Medicine, Osaka University.

    Techniques: Injection, Transgenic Assay, Mouse Assay, Isolation, Staining