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Hypoxia/reoxygenation (H/R)‐induced injury in human ventricular cardiac organoids. (A) Modelling process of H/R in ventricular cardiac organoids. (B) Contour plots of flow cytometry analysis for apoptotic cells in control and H/R groups by Annexin V‐FITC/PI‐PE staining. (C) Proportions of early and late apoptosis cells in flow cytometry analysis for control and H/R groups, n = 5 independent replicates. (D) Distribution of TNNT2 (red), TUNEL (green) and DAPI (blue) in control and H/R groups. Scale bars (left), 200 μm. Scale bars (right), 20 μm. (E) MDA concentration in control and H/R groups, n = 7 independent replicates. (F) Relative cytotoxicity of lactate dehydrogenase (LDH) in control and H/R groups, n = 6 independent replicates. (G) Distribution of CMs (TNNT2), ECs (CD31), and fibroblasts (VIM) in control and H/R groups. Scale bars (left), 200 μm. Scale bars (right), 20 μm. (H) Bright‐field microscopy images and schematic representation of <t>MEA‐measured</t> contractility and Beat Amplitude values ( n = 23 electrodes per group) in control and H/R groups. (I) Representative calcium images at active and rest stages in control and H/R groups and the synchronized contraction was shown by calcium flux frequency. Results were expressed as F/F0 (F0 refers to baseline intensity). Scale bar, 200 μm. H/R indicates hypoxia/reoxygenation and VIM indicates Vimentin. Bar and dot plot graphs show mean ± SD. Statistical significance was assessed by unpaired t ‐test (*** p < 0.001).
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Hypoxia/reoxygenation (H/R)‐induced injury in human ventricular cardiac organoids. (A) Modelling process of H/R in ventricular cardiac organoids. (B) Contour plots of flow cytometry analysis for apoptotic cells in control and H/R groups by Annexin V‐FITC/PI‐PE staining. (C) Proportions of early and late apoptosis cells in flow cytometry analysis for control and H/R groups, n = 5 independent replicates. (D) Distribution of TNNT2 (red), TUNEL (green) and DAPI (blue) in control and H/R groups. Scale bars (left), 200 μm. Scale bars (right), 20 μm. (E) MDA concentration in control and H/R groups, n = 7 independent replicates. (F) Relative cytotoxicity of lactate dehydrogenase (LDH) in control and H/R groups, n = 6 independent replicates. (G) Distribution of CMs (TNNT2), ECs (CD31), and fibroblasts (VIM) in control and H/R groups. Scale bars (left), 200 μm. Scale bars (right), 20 μm. (H) Bright‐field microscopy images and schematic representation of <t>MEA‐measured</t> contractility and Beat Amplitude values ( n = 23 electrodes per group) in control and H/R groups. (I) Representative calcium images at active and rest stages in control and H/R groups and the synchronized contraction was shown by calcium flux frequency. Results were expressed as F/F0 (F0 refers to baseline intensity). Scale bar, 200 μm. H/R indicates hypoxia/reoxygenation and VIM indicates Vimentin. Bar and dot plot graphs show mean ± SD. Statistical significance was assessed by unpaired t ‐test (*** p < 0.001).
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Hypoxia/reoxygenation (H/R)‐induced injury in human ventricular cardiac organoids. (A) Modelling process of H/R in ventricular cardiac organoids. (B) Contour plots of flow cytometry analysis for apoptotic cells in control and H/R groups by Annexin V‐FITC/PI‐PE staining. (C) Proportions of early and late apoptosis cells in flow cytometry analysis for control and H/R groups, n = 5 independent replicates. (D) Distribution of TNNT2 (red), TUNEL (green) and DAPI (blue) in control and H/R groups. Scale bars (left), 200 μm. Scale bars (right), 20 μm. (E) MDA concentration in control and H/R groups, n = 7 independent replicates. (F) Relative cytotoxicity of lactate dehydrogenase (LDH) in control and H/R groups, n = 6 independent replicates. (G) Distribution of CMs (TNNT2), ECs (CD31), and fibroblasts (VIM) in control and H/R groups. Scale bars (left), 200 μm. Scale bars (right), 20 μm. (H) Bright‐field microscopy images and schematic representation of <t>MEA‐measured</t> contractility and Beat Amplitude values ( n = 23 electrodes per group) in control and H/R groups. (I) Representative calcium images at active and rest stages in control and H/R groups and the synchronized contraction was shown by calcium flux frequency. Results were expressed as F/F0 (F0 refers to baseline intensity). Scale bar, 200 μm. H/R indicates hypoxia/reoxygenation and VIM indicates Vimentin. Bar and dot plot graphs show mean ± SD. Statistical significance was assessed by unpaired t ‐test (*** p < 0.001).
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Hypoxia/reoxygenation (H/R)‐induced injury in human ventricular cardiac organoids. (A) Modelling process of H/R in ventricular cardiac organoids. (B) Contour plots of flow cytometry analysis for apoptotic cells in control and H/R groups by Annexin V‐FITC/PI‐PE staining. (C) Proportions of early and late apoptosis cells in flow cytometry analysis for control and H/R groups, n = 5 independent replicates. (D) Distribution of TNNT2 (red), TUNEL (green) and DAPI (blue) in control and H/R groups. Scale bars (left), 200 μm. Scale bars (right), 20 μm. (E) MDA concentration in control and H/R groups, n = 7 independent replicates. (F) Relative cytotoxicity of lactate dehydrogenase (LDH) in control and H/R groups, n = 6 independent replicates. (G) Distribution of CMs (TNNT2), ECs (CD31), and fibroblasts (VIM) in control and H/R groups. Scale bars (left), 200 μm. Scale bars (right), 20 μm. (H) Bright‐field microscopy images and schematic representation of <t>MEA‐measured</t> contractility and Beat Amplitude values ( n = 23 electrodes per group) in control and H/R groups. (I) Representative calcium images at active and rest stages in control and H/R groups and the synchronized contraction was shown by calcium flux frequency. Results were expressed as F/F0 (F0 refers to baseline intensity). Scale bar, 200 μm. H/R indicates hypoxia/reoxygenation and VIM indicates Vimentin. Bar and dot plot graphs show mean ± SD. Statistical significance was assessed by unpaired t ‐test (*** p < 0.001).
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Hypoxia/reoxygenation (H/R)‐induced injury in human ventricular cardiac organoids. (A) Modelling process of H/R in ventricular cardiac organoids. (B) Contour plots of flow cytometry analysis for apoptotic cells in control and H/R groups by Annexin V‐FITC/PI‐PE staining. (C) Proportions of early and late apoptosis cells in flow cytometry analysis for control and H/R groups, n = 5 independent replicates. (D) Distribution of TNNT2 (red), TUNEL (green) and DAPI (blue) in control and H/R groups. Scale bars (left), 200 μm. Scale bars (right), 20 μm. (E) MDA concentration in control and H/R groups, n = 7 independent replicates. (F) Relative cytotoxicity of lactate dehydrogenase (LDH) in control and H/R groups, n = 6 independent replicates. (G) Distribution of CMs (TNNT2), ECs (CD31), and fibroblasts (VIM) in control and H/R groups. Scale bars (left), 200 μm. Scale bars (right), 20 μm. (H) Bright‐field microscopy images and schematic representation of MEA‐measured contractility and Beat Amplitude values ( n = 23 electrodes per group) in control and H/R groups. (I) Representative calcium images at active and rest stages in control and H/R groups and the synchronized contraction was shown by calcium flux frequency. Results were expressed as F/F0 (F0 refers to baseline intensity). Scale bar, 200 μm. H/R indicates hypoxia/reoxygenation and VIM indicates Vimentin. Bar and dot plot graphs show mean ± SD. Statistical significance was assessed by unpaired t ‐test (*** p < 0.001).

Journal: Cell Proliferation

Article Title: Modelling myocardial ischemia/reperfusion injury with inflammatory response in human ventricular cardiac organoids

doi: 10.1111/cpr.13762

Figure Lengend Snippet: Hypoxia/reoxygenation (H/R)‐induced injury in human ventricular cardiac organoids. (A) Modelling process of H/R in ventricular cardiac organoids. (B) Contour plots of flow cytometry analysis for apoptotic cells in control and H/R groups by Annexin V‐FITC/PI‐PE staining. (C) Proportions of early and late apoptosis cells in flow cytometry analysis for control and H/R groups, n = 5 independent replicates. (D) Distribution of TNNT2 (red), TUNEL (green) and DAPI (blue) in control and H/R groups. Scale bars (left), 200 μm. Scale bars (right), 20 μm. (E) MDA concentration in control and H/R groups, n = 7 independent replicates. (F) Relative cytotoxicity of lactate dehydrogenase (LDH) in control and H/R groups, n = 6 independent replicates. (G) Distribution of CMs (TNNT2), ECs (CD31), and fibroblasts (VIM) in control and H/R groups. Scale bars (left), 200 μm. Scale bars (right), 20 μm. (H) Bright‐field microscopy images and schematic representation of MEA‐measured contractility and Beat Amplitude values ( n = 23 electrodes per group) in control and H/R groups. (I) Representative calcium images at active and rest stages in control and H/R groups and the synchronized contraction was shown by calcium flux frequency. Results were expressed as F/F0 (F0 refers to baseline intensity). Scale bar, 200 μm. H/R indicates hypoxia/reoxygenation and VIM indicates Vimentin. Bar and dot plot graphs show mean ± SD. Statistical significance was assessed by unpaired t ‐test (*** p < 0.001).

Article Snippet: The multi‐electrode array (MEA) plate was transferred to the MEA device, Maestro Edge version (Axion BioSystems Inc.; Atlanta, GA, USA) for measuring the spike amplitude and beat amplitude of field potential under spontaneous beating without electrical stimulation.

Techniques: Flow Cytometry, Control, Staining, TUNEL Assay, Concentration Assay, Microscopy