Journal: Stem Cell Reports

Article Title: Aberrant α-Adrenergic Hypertrophic Response in Cardiomyocytes from Human Induced Pluripotent Cells

doi: 10.1016/j.stemcr.2014.09.002

Figure Lengend Snippet: Distinct Responses of Cardiomyocyte Derived from hESC and hiPSC Lines to Phenylephrine (A) Representative immunofluorescence confocal image and 3D rendering of a confocal image stack showing differentiated hiPSC-CMs stained positive for cardiac-specific myosin heavy chain α/β (green) and ANF (red) at 30 days after differentiation. Nuclei are stained with DAPI (blue). White scale bars represent 20 μm. Cardiomyocytes differentiated from various hESC (H7, HUES7, and SHEF3) and hiPSC (LQT2, LQT2-PAT, CDI, and ReproCell) lines showed comparable morphology after plating onto 0.5% gelatine. The hESC-CMs and hiPSC-CMs were treated with phenylephrine (PE) (10 μM, 48 hr) at 30 days after differentiation. (B and C) Bar graphs showing fold change in 2D cell area (B) and ANF mRNA levels (C) in cells treated with pPE. Results are shown as fold changes versus control group. n > 100 MHC-positive cells analyzed per well (mean ± SEM), from two to ten biological replicates. ∗ p < 0.05, ∗∗∗ p < 0.001 versus control groups; Student’s t test. See also and .

Article Snippet: Increases in cell size somewhat larger than this have been reported for iCell hiPSC-CMs with ET-1 (24% versus our 10%) , but it is notable that the stronger BNP signal was chosen for the final hypertrophy assay in that study, which would be consistent with our findings.

Techniques: Derivative Assay, Immunofluorescence, Staining

Journal: Stem Cell Reports

Article Title: Aberrant α-Adrenergic Hypertrophic Response in Cardiomyocytes from Human Induced Pluripotent Cells

doi: 10.1016/j.stemcr.2014.09.002

Figure Lengend Snippet: Upregulation of ADRA1B Expression during Differentiation of hESCs and hiPSCs (A) Quantitative RT-PCR data on ADRA1B expressions in undifferentiated hESCs, hiPSCs, hiPSC-CMs, hESC-CMs, and adult myocytes isolated from the left ventricle, adult heart, and fetal heart and fibroblasts. Undifferentiated cells are shown as gray, differentiated cardiomyocytes as red, differentiated fibroblasts as blue, adult cardiomyocytes as dark red, and adult fibroblasts as dark blue bars. Data are expressed as relative changes versus DMSO-treated differentiating control hESCs or hiPSCs; mean ± SEM, ∗∗∗ p < 0.001 versus undifferentiated group; Student’s t test. Samples are from three biological replicates. (B) The diagram in Figure 2 shows the experimental scheme of differentiation and reprogramming. Bar graphs showing ADRA1B mRNA in HUES7-hESC-, HUES7-hiPSC-, and HUES7-hiPSC-derived cardiomyocytes or fibroblasts, adult skin fibroblasts, and hiPSC- and hiPSC-derived cardiomyocytes from a patient with LQT2 syndrome and a healthy relative (LQT2-PAT) are presented. Undifferentiated cells are shown as gray, differentiated cardiomyocytes as red, differentiated fibroblasts as blue, adult cardiomyocytes as dark red, and adult fibroblasts as dark blue bars. Results are shown as fold changes versus undifferentiated hESC group; mean ± SEM. Samples were measured in triplicate from two biological replicates.

Article Snippet: Increases in cell size somewhat larger than this have been reported for iCell hiPSC-CMs with ET-1 (24% versus our 10%) , but it is notable that the stronger BNP signal was chosen for the final hypertrophy assay in that study, which would be consistent with our findings.

Techniques: Expressing, Quantitative RT-PCR, Isolation, Derivative Assay

Journal: Stem Cell Reports

Article Title: Aberrant α-Adrenergic Hypertrophic Response in Cardiomyocytes from Human Induced Pluripotent Cells

doi: 10.1016/j.stemcr.2014.09.002

Figure Lengend Snippet: Overexpression of ADRA1A in hiPSC-CM (A) Localization of ADRA1A in hiPSC-CM (iCell) transfected with ADRA1A-eYFP construct. White scale bar represents 20 μm. (B–D) Bar graphs showing ADRA1A mRNA levels (B), cell size (C), and ANF mRNA levels (D) in hiPSC-CMs transfected with ADRA1A without or in the presence of PE after 48 hr; mean ± SEM; six biological replicates. ∗∗∗ p < 0.001 versus control group; one-way ANOVA with Tukey’s post hoc test.

Article Snippet: Increases in cell size somewhat larger than this have been reported for iCell hiPSC-CMs with ET-1 (24% versus our 10%) , but it is notable that the stronger BNP signal was chosen for the final hypertrophy assay in that study, which would be consistent with our findings.

Techniques: Over Expression, Transfection, Construct

Journal: Stem Cell Reports

Article Title: Aberrant α-Adrenergic Hypertrophic Response in Cardiomyocytes from Human Induced Pluripotent Cells

doi: 10.1016/j.stemcr.2014.09.002

Figure Lengend Snippet: Distinct Expressions of Downstream G Protein during Differentiation from hESCs and hiPSCs toward hESC-CMs and hiPSC-CMs (A–C) mRNA for Gq (A), Gβ1 (B), and Gγ2 (C) proteins were all detectable in various lines of undifferentiated hESC (light gray bars) and hiPSC (dark gray bars) cultures, fetal hearts, and fibroblasts and adult ventricular cardiomyocytes from heart failure patients (black bars). Results are shown as fold changes versus the undifferentiated hESC (HUES7) group (mean ± SEM; three biological replicates). (D–F) Changes in mRNA levels for Gq (D), Gβ1 (E), and Gγ2 (F) during differentiation of hESCs of hiPSCs are also presented. Results are shown as fold changes versus the respective undifferentiated hESC or hiPSC line. Samples were measured in triplicate from three biological replicates. ∗ p < 0.05, ∗∗∗ p < 0.001; Student’s t test.

Article Snippet: Increases in cell size somewhat larger than this have been reported for iCell hiPSC-CMs with ET-1 (24% versus our 10%) , but it is notable that the stronger BNP signal was chosen for the final hypertrophy assay in that study, which would be consistent with our findings.

Techniques:

Journal: Stem Cell Reports

Article Title: Aberrant α-Adrenergic Hypertrophic Response in Cardiomyocytes from Human Induced Pluripotent Cells

doi: 10.1016/j.stemcr.2014.09.002

Figure Lengend Snippet: Phosphokinase Proteome and Kinase Inhibitor Analyses of PE-Induced Signaling in hESC-CMs and hiPSC-CMs (A) Relative changes in phosphoprotein levels in response to PE (10 μM, 48 hr) in hESC-CMs (H7) and hiPSC-CMs (iCell). (B) Schematic diagram of Ingenuity Pathway Analysis-mapped mechanistic interactions of active epidermal growth factor receptor kinase, STAT family members, and GSK3β/ β-catenin and Src kinase pathways. Arrows between nodes represent direct (solid lines) and indirect (dashed lines) interactions between molecules as supported by information in the Ingenuity Pathway Knowledge Base. (C) Assessment of kinase inhibitors on hypertrophy. Human ESC-CMs (H7) and hiPSC-CMs (iCell and ReproCell) were treated with PE in the presence of kinase inhibitors. Bar graphs show changes in response to selected kinase inhibitors on cell area by automated high-content microscopy. Robust Z score was computed and visualized. (D–F) Representative image and quantitation of nuclear translocation of STAT3 (D and E) and cell area (F) in hiPSC-CMs treated with PE in the presence of interleukin-6 (100 ng/ml) in hiPSC-CMs. White scale bar represents 20 μm. (G) Cell area in hiPSC-CMs treated with PE in the presence of combined inhibition of GSK3β/EGFRK/CAMKII/src/PDGFRK (1 μM each); mean ± SEM; four biological replicates; one-way ANOVA with Tukey’s post hoc test. See also and .

Article Snippet: Increases in cell size somewhat larger than this have been reported for iCell hiPSC-CMs with ET-1 (24% versus our 10%) , but it is notable that the stronger BNP signal was chosen for the final hypertrophy assay in that study, which would be consistent with our findings.

Techniques: Microscopy, Quantitation Assay, Translocation Assay, Inhibition

Journal: Stem Cell Reports

Article Title: Aberrant α-Adrenergic Hypertrophic Response in Cardiomyocytes from Human Induced Pluripotent Cells

doi: 10.1016/j.stemcr.2014.09.002

Figure Lengend Snippet: Schematic Summary of Potential Differences in Adrenergic Receptor-Driven Regulation of Cell Growth and Hypertrophy in hiPSC-CMs and hESC-CMs Red is used to represent active/upregulated components and gray for relatively inactive or downregulated components.

Article Snippet: Increases in cell size somewhat larger than this have been reported for iCell hiPSC-CMs with ET-1 (24% versus our 10%) , but it is notable that the stronger BNP signal was chosen for the final hypertrophy assay in that study, which would be consistent with our findings.

Techniques:

Journal: Discover Applied Sciences

Article Title: Identifying and establishing the critical elements of a human cardiac in-vitro model for studying type-II diabetes

doi: 10.1007/s42452-025-07442-y

Figure Lengend Snippet: Dose dependent effects of AGEs on AC16 cardiomyocytes. The varying doses applied included 100 µg, 250 and 500 µg for 24 h after which the cells were washed and replenished with complete growth medium. A live cell staining assay was performed to assess the effects of the varying doses of AGEs shown above. Scale bar indicates 100 μm

Article Snippet: Building on existing work, in this study, we exposed human cardiomyocytes, AC16, and human induced pluripotent stem cell-derived cardiomyocytes (iCell cardiomyocytes) [ ] to an optimized dose of AGEs, or hyperglycemia to compare their effects on cardiac function.

Techniques: Staining

Journal: Discover Applied Sciences

Article Title: Identifying and establishing the critical elements of a human cardiac in-vitro model for studying type-II diabetes

doi: 10.1007/s42452-025-07442-y

Figure Lengend Snippet: Expression Fold Change for gene expression of Cx-43 (encoded by GJA1) in AC16 cardiomyocytes after 48 h of AGEs exposure and post washouts. Data is represented as a normalized relative fold change to control. p-values between control and AGEs or AGEs washouts were significant (0.02) however in between AGEs and AGEs washout no significance was noted

Article Snippet: Building on existing work, in this study, we exposed human cardiomyocytes, AC16, and human induced pluripotent stem cell-derived cardiomyocytes (iCell cardiomyocytes) [ ] to an optimized dose of AGEs, or hyperglycemia to compare their effects on cardiac function.

Techniques: Expressing, Gene Expression, Control

Journal: Discover Applied Sciences

Article Title: Identifying and establishing the critical elements of a human cardiac in-vitro model for studying type-II diabetes

doi: 10.1007/s42452-025-07442-y

Figure Lengend Snippet: AC16 Cardiomyocytes Cardiac Myosin Heavy Chain (MHC) immunostaining for different experimental conditions: ( A ) Control, ( B ) AGEs, ( C ) Glucose Shock (50 mM), ( D ) AGEs followed by Glucose Shock, ( E ) Inhibitor followed by AGEs, ( F ) Inhibitor followed by AGEs and Glucose Shock. ( G ) Shows the expression levels of cardiac MHC calculated, data is represented as a normalized mean intensity to control. * Represents p < 0.05

Article Snippet: Building on existing work, in this study, we exposed human cardiomyocytes, AC16, and human induced pluripotent stem cell-derived cardiomyocytes (iCell cardiomyocytes) [ ] to an optimized dose of AGEs, or hyperglycemia to compare their effects on cardiac function.

Techniques: Immunostaining, Control, Expressing

Journal: Discover Applied Sciences

Article Title: Identifying and establishing the critical elements of a human cardiac in-vitro model for studying type-II diabetes

doi: 10.1007/s42452-025-07442-y

Figure Lengend Snippet: AC16 Cardiomyocytes Connexin 43 (Cx-43) immunostaining for different experimental conditions: ( A ) Control, ( B ) AGEs, ( C ) Glucose Shock (50 mM), ( D ) AGEs followed by Glucose Shock, ( E ) Inhibitor followed by AGEs, ( F ) Inhibitor followed by AGEs and Glucose Shock. ( G ) Shows the expression levels of Cx-43 calculated, data is represented as a normalized relative mean intensity to control. * Represents p < 0.05

Article Snippet: Building on existing work, in this study, we exposed human cardiomyocytes, AC16, and human induced pluripotent stem cell-derived cardiomyocytes (iCell cardiomyocytes) [ ] to an optimized dose of AGEs, or hyperglycemia to compare their effects on cardiac function.

Techniques: Immunostaining, Control, Expressing

Journal: Discover Applied Sciences

Article Title: Identifying and establishing the critical elements of a human cardiac in-vitro model for studying type-II diabetes

doi: 10.1007/s42452-025-07442-y

Figure Lengend Snippet: AC16 Cardiomyocytes RAGE expression for different experimental conditions: ( A ) Control, ( B ) AGEs (500 µg/ml), ( C ) Glucose Shock (50 mM), ( D ) AGEs followed by Glucose Shock, ( E ) Shows the expression levels of RAGE calculated, data is represented as a normalized relative mean intensity to DAPI. * Represents p < 0.05

Article Snippet: Building on existing work, in this study, we exposed human cardiomyocytes, AC16, and human induced pluripotent stem cell-derived cardiomyocytes (iCell cardiomyocytes) [ ] to an optimized dose of AGEs, or hyperglycemia to compare their effects on cardiac function.

Techniques: Expressing, Control

Journal: Discover Applied Sciences

Article Title: Identifying and establishing the critical elements of a human cardiac in-vitro model for studying type-II diabetes

doi: 10.1007/s42452-025-07442-y

Figure Lengend Snippet: Expression Fold Change for gene expression of Cx-43 (encoded by GJA1) in AC16 cardiomyocytes under different experimental conditions after 24 h of exposure. Data is represented as a normalized relative fold change to control. * Represents p < 0.05

Article Snippet: Building on existing work, in this study, we exposed human cardiomyocytes, AC16, and human induced pluripotent stem cell-derived cardiomyocytes (iCell cardiomyocytes) [ ] to an optimized dose of AGEs, or hyperglycemia to compare their effects on cardiac function.

Techniques: Expressing, Gene Expression, Control

Journal: Discover Applied Sciences

Article Title: Identifying and establishing the critical elements of a human cardiac in-vitro model for studying type-II diabetes

doi: 10.1007/s42452-025-07442-y

Figure Lengend Snippet: Dose dependent effects of AGEs on iCell Cardiomyocytes. The varying doses applied included ( A ) control, ( B ) 100 µg/ml, ( C ) 250 µg/ml, and ( D ) 500 µg/ml for 30 min after which the cells were washed and replenished with complete growth medium. ( E ) Shows the percentage cell viability measured from the live dead assay performed to assess the effects of the varying doses of AGEs.

Article Snippet: Building on existing work, in this study, we exposed human cardiomyocytes, AC16, and human induced pluripotent stem cell-derived cardiomyocytes (iCell cardiomyocytes) [ ] to an optimized dose of AGEs, or hyperglycemia to compare their effects on cardiac function.

Techniques: Control, Live Dead Assay

Journal: Discover Applied Sciences

Article Title: Identifying and establishing the critical elements of a human cardiac in-vitro model for studying type-II diabetes

doi: 10.1007/s42452-025-07442-y

Figure Lengend Snippet: Beat Rate heat maps and waveforms studied in relation with dose dependent effects of AGEs on iCell Cardiomyocytes. Image shows applied AGEs at a 100 µg/ml concentration for 30 min after which their contractility characterization was performed. Controls included wells with cells that did not receive any treatments and were washed and replenished with complete growth medium. The figure labels are as follows: ( A ) Control (Scale: 0 to 76), ( B ) AGEs (Scale: 0 to 56), ( C ) AGEs + Glucose Shock (Scale: 0 to 70), ( D ) AGEs Inhibitor + AGEs (Scale: 0 to 56), and ( E ) AGEs Inhibitor + AGEs 100 µg/ml + Glucose Shock (Scale: 0 to 70)

Article Snippet: Building on existing work, in this study, we exposed human cardiomyocytes, AC16, and human induced pluripotent stem cell-derived cardiomyocytes (iCell cardiomyocytes) [ ] to an optimized dose of AGEs, or hyperglycemia to compare their effects on cardiac function.

Techniques: Concentration Assay, Control

Journal: Discover Applied Sciences

Article Title: Identifying and establishing the critical elements of a human cardiac in-vitro model for studying type-II diabetes

doi: 10.1007/s42452-025-07442-y

Figure Lengend Snippet: Beat Rate heat maps and waveforms studied in relation with dose dependent effects of AGEs on iCell Cardiomyocytes. Image shows applied AGEs at a 250 µg/ml concentration for 30 min after which their contractility characterization was performed. Controls included wells with cells that did not receive any treatments and were washed and replenished with complete growth medium. The figure labels are as follows: ( A ) Control (Scale: 0 to 76), ( B ) AGEs (Scale: 0 to 42), ( C ) AGEs + Glucose Shock (Scale: 0 to 71), ( D ) AGEs Inhibitor + AGEs (Scale: 0 to 42), and ( E ) AGEs Inhibitor + AGEs 250 µg/ml + Glucose Shock (Scale: 0 to 71)

Article Snippet: Building on existing work, in this study, we exposed human cardiomyocytes, AC16, and human induced pluripotent stem cell-derived cardiomyocytes (iCell cardiomyocytes) [ ] to an optimized dose of AGEs, or hyperglycemia to compare their effects on cardiac function.

Techniques: Concentration Assay, Control

Journal: Discover Applied Sciences

Article Title: Identifying and establishing the critical elements of a human cardiac in-vitro model for studying type-II diabetes

doi: 10.1007/s42452-025-07442-y

Figure Lengend Snippet: Beats per minute (BPM) or cardiac contractile behavior studied in relation with dose dependent effects of AGES on iCell Cardiomyocytes. The varying doses applied included 100 and 250 µg/ml for 30 min after which they were recorded. Controls included wells with cells that did not receive any treatments and were washed and replenished with complete growth medium. * Represents p < 0.05

Article Snippet: Building on existing work, in this study, we exposed human cardiomyocytes, AC16, and human induced pluripotent stem cell-derived cardiomyocytes (iCell cardiomyocytes) [ ] to an optimized dose of AGEs, or hyperglycemia to compare their effects on cardiac function.

Techniques:

Journal: Discover Applied Sciences

Article Title: Identifying and establishing the critical elements of a human cardiac in-vitro model for studying type-II diabetes

doi: 10.1007/s42452-025-07442-y

Figure Lengend Snippet: iCell Cardiomyocytes control wells (no treatments): ( A ) Brightfield, ( B ) immunostaining and ( C ) RT-PCR of Cardiac MHC (encoded by MYH7)

Article Snippet: Building on existing work, in this study, we exposed human cardiomyocytes, AC16, and human induced pluripotent stem cell-derived cardiomyocytes (iCell cardiomyocytes) [ ] to an optimized dose of AGEs, or hyperglycemia to compare their effects on cardiac function.

Techniques: Control, Immunostaining, Reverse Transcription Polymerase Chain Reaction

Journal: Scientific Reports

Article Title: Human perinatal stem cell derived extracellular matrix enables rapid maturation of hiPSC-CM structural and functional phenotypes

doi: 10.1038/s41598-020-76052-y

Figure Lengend Snippet: Effects of ECM on electrophysiology and drug-response of hiPSC-CMs. ( A , B ) hiPSC-CMs cultured on Matrigel, CELLvo or Matrix Plus were treated E-4031 (500 nM). Monolayers plated on Matrix Plus did not have a significant change in beat rate (n = 6, Δ beat rate = + 0.22 Hz; p = 0.343), but had a significant increase in APD80 (n = 6, Δ APD80 = + 1,227.5 ms; p = 0.002) formation of rotors (TdPs) in 100% tested; whilst cells cultured on Matrigel presented a significant decrease in beat rate and APD80 prolongation and EADs that transitioned to TdPs in just 16% tested. Syncytia cultured on CELLvo had APD80 prolongation, but no incidence of early-afterdepolarizations (EADs) or TdPs. ( C , D ) Treatment with 100 µM ranolazine, a safe drug did not change spontaneous beat rate in hiPSC-CMs cultured on Matrigel (n = 6, Δ beat rate = 0 Hz; p = 0.485), CELLvo (n = 6, Δ beat rate = − 0.1 Hz; p = 0.109) or Matrix Plus (n = 6, Δ beat rate = − 0.02 Hz; p = 0.817); Additionally, after ranolazine treatment cardiomyocyte syncytia cultured on Matrix Plus presented APD80 prolongation (n = 6, Δ APD80 = 66.3 ms; p = 0.009) without presence of EADs or TdPs, similar results were obtained with syncytia cultured on CELLvo (n = 6, Δ APD80 = 52.4 ms; p = 0.014) and Matrigel (n = 6, Δ APD80 = 77.9 ms; p < 0.001). ( E ) Microelectrode recording of spontaneous action potentials was done to quantify membrane potentials. Action potential upstroke velocity was greater on cells cultured on Matrix Plus in comparison to Matrigel (p = 0.002). Conduction velocity was significantly higher on Matrigel than Matrix Plus (p = 0.01).

Article Snippet: In these experiments cryopreserved hiPSC-CMs (Cellular Dynamics International, iCell ) were plated as electrically and mechanically connected monolayers on one of three ECMs: 1.

Techniques: Cell Culture

Journal: Scientific Reports

Article Title: Human perinatal stem cell derived extracellular matrix enables rapid maturation of hiPSC-CM structural and functional phenotypes

doi: 10.1038/s41598-020-76052-y

Figure Lengend Snippet: Matrix Plus high-throughput screening assay allows observation of arrhythmias. ( A ) Ninety-six well plates coated with Matrix Plus were submitted to optical mapping for voltage changes and presented a baseline beat rate variation below 6%. ( B ) hiPSC-CMs cultured on Matrigel were treated with drugs classified as high, intermediate and low risk at 10 × ETPC (historical data) were compared to hiPSC-CMs cultured on Matrix Plus and treated with a matched set of drugs at the same concentration. Whilst most of drugs were not able to induce arrhythmias at 10 × ETPC in syncytia cultured on Matrigel, there was a progressive increase in the number of arrhythmias associated with increase in drug risk in the syncytia cultured on Matrix Plus. ( C ) Syncytia culture on Matrix plus were treated with 25 different drugs classifies as high, intermediate and low risk and there was a proportionated increase in the number of arrhythmias corresponding directly to the risk group; nevertheless, increase in arrhythmias was not observed in syncytia cultured on Matrigel.

Article Snippet: In these experiments cryopreserved hiPSC-CMs (Cellular Dynamics International, iCell ) were plated as electrically and mechanically connected monolayers on one of three ECMs: 1.

Techniques: High Throughput Screening Assay, Screening Assay, Cell Culture, Concentration Assay

Journal: Scientific Reports

Article Title: Human perinatal stem cell derived extracellular matrix enables rapid maturation of hiPSC-CM structural and functional phenotypes

doi: 10.1038/s41598-020-76052-y

Figure Lengend Snippet: Matrix Plus promotes structural maturation of hiPSC-CMs in 7 days. ( A , C ) Bright-field imaging of hiPSC-CM plated in low density shows that Matrix Plus enables attachment comparable to Matrigel, with morphological differences despite using the same batch of cells for each condition. ( B , D ) Matrix plus also permits the formation of higher density functional syncytia with preservation of morphology changes that were not observed on Matrigel. ( E , F ) Sarcomere staining using α-actinin shows morphological maturation of hiPSC-CMs induced by Matrix Plus.

Article Snippet: In these experiments cryopreserved hiPSC-CMs (Cellular Dynamics International, iCell ) were plated as electrically and mechanically connected monolayers on one of three ECMs: 1.

Techniques: Imaging, Functional Assay, Preserving, Staining

Journal: Scientific Reports

Article Title: Human perinatal stem cell derived extracellular matrix enables rapid maturation of hiPSC-CM structural and functional phenotypes

doi: 10.1038/s41598-020-76052-y

Figure Lengend Snippet: Matrix plus induces morphology changes compatible with adult-like phenotype. ( A – C ) TnI, cTnT and α-actinin immunostaining collectively indicate Matrix Plus induces rod-shape morphology with organization of sarcomeres parallel to the long axis of the cardiomyocyte. ( C ) hiPSC-CM circularity index was significantly higher on Matrigel (n = 52; circularity index: 0.71 ± 0.02) than on Matrix Plus (n = 88; circularity index: 0.38 ± 0.01; p < 0.0001). ( D ) Circularity index was calculated using fluorescent images of sarcomere staining. The circularity index for hiPSC-CMs was greater on Matrigel than on Matrix Plus (n = 52, 88 for Matrigel and Matrix Plus respectively, P < 0.0001, unpaired t-test).

Article Snippet: In these experiments cryopreserved hiPSC-CMs (Cellular Dynamics International, iCell ) were plated as electrically and mechanically connected monolayers on one of three ECMs: 1.

Techniques: Immunostaining, Staining

Journal: Scientific Reports

Article Title: Human perinatal stem cell derived extracellular matrix enables rapid maturation of hiPSC-CM structural and functional phenotypes

doi: 10.1038/s41598-020-76052-y

Figure Lengend Snippet: Matrix Plus promotes higher mitochondrial membrane potential. ( A – F ) hiPSC-CMs cultured on Matrigel (n = 30, mitrotracker intensity/cell: 221 ± 14.75 a.u.; p < 0.0001) have a lower mitochrondrial content in comparison to cardiomyocytes cultured on Matrix Plus (n = 40, mitotracker intensity/cell: 569 ± 34.9a.u.) and murine adult ventricular cardiomyocytes (n = 56, mitotracker intensity/cell: 859 ± 22.3a.u.).( G – M ) Mitochondria functional information obtained with JC-1 also indicates that cardiomyocytes cultured on Matrix Plus have higher mitochondrial membrane potential than cardiomyocytes cultured on Matrigel (p < 0.0001).

Article Snippet: In these experiments cryopreserved hiPSC-CMs (Cellular Dynamics International, iCell ) were plated as electrically and mechanically connected monolayers on one of three ECMs: 1.

Techniques: Cell Culture, Functional Assay

Journal: Scientific Reports

Article Title: Emergent synchronous beating behavior in spontaneous beating cardiomyocyte clusters

doi: 10.1038/s41598-021-91466-y

Figure Lengend Snippet: Formation of mouse primary cardiomyocyte clusters in agarose-coated wells. ( a ) Schematic drawing of the conventional dish cultivation of cardiomyocytes. The dispersed cells were cultured on the bottom of a 35-mm non-agarose-coated dish. After spread of the 2.0 mL of \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$5.0\times 10^{4}\, {\rm cells/mL}$$\end{document} 5.0 × 10 4 cells / mL isolated single cardiomyocytes, the cells attached on the bottom of the 35-mm cultivation dish dispersedly. The cells started to beat 2–3 days after cultivation started. ( b ) A micrograph of dispersed cardiomyocytes in a 35-mm non-agarose-coated dish. ( c ) Schematic drawing of the cultivation of dispersed cells in a 35-mm agarose-coated dish. After spread of the 2.0 mL of \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$5.0\times 10^{4}\, {\rm cells/mL}$$\end{document} 5.0 × 10 4 cells / mL isolated single cardiomyocytes, the cells dispersed on the bottom of the agarose layer in the agarose-coated 35-mm cultivation dish. Even after 2–3 days of cultivation, the cells remained isolated with a round shape, and no clusters formed on the bottom. ( d ) A micrograph of cardiomyocytes in an agarose-coated 35-mm cultivation dish. ( e ) Schematic drawing of the cultivation of dispersed cells in a 15.5-mm agarose-coated cultivation well (in a 24-well cultivation plate). After spread of the 1.0 mL of \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$5\times 10^{4}\hbox { cells/mL}$$\end{document} 5 × 10 4 cells/mL isolated single cardiomyocytes, dispersed cells gathered and formed small clusters; finally, they gathered into a single large cluster in the 15.5-mm agarose-coated cultivation well. ( f ) A micrograph of a cardiomyocyte cluster in a 15.5-mm agarose-coated cultivation well.

Article Snippet: Human embryonic stem cell-derived cardiomyocytes (hES) (hES-CMCTM002, hES cell line SA002) were purchased from Cellectis (Gothenburg, Sweden) , .

Techniques: Cell Culture, Isolation

Journal: Scientific Reports

Article Title: Emergent synchronous beating behavior in spontaneous beating cardiomyocyte clusters

doi: 10.1038/s41598-021-91466-y

Figure Lengend Snippet: Micrographs of single cells and clusters of mouse primary and hES-derived cardiomyocytes. ( a ) Mouse primary cardiomyocytes (primary) in a 35-mm non-agarose-coated dish (single cell), ( b ) primary cells in a 24-well agarose-coated plate (cluster), ( c ) hES cardiomyocytes in a 35-mm non-agarose-coated dish (single cell), and ( d ) hES in a 24-well agarose-coated plate (cluster).

Article Snippet: Human embryonic stem cell-derived cardiomyocytes (hES) (hES-CMCTM002, hES cell line SA002) were purchased from Cellectis (Gothenburg, Sweden) , .

Techniques: Derivative Assay

Journal: Scientific Reports

Article Title: Emergent synchronous beating behavior in spontaneous beating cardiomyocyte clusters

doi: 10.1038/s41598-021-91466-y

Figure Lengend Snippet: Analysis of interbeat interval (IBI) distribution of single and clustered mouse primary and hES cardiomyocytes. ( a )–( d ): Method of measuring interbeat interval (IBI) of single cardiomyocytes and clusters. Temporal change of luminance in the red square area for single cell ( a ) and cluster ( c ) caused by their beating was recorded, as shown in the time-course intensity profiles ( b ) and ( d ), respectively. IBIs of their beating were acquired from the time intervals between two neighboring peaks in the time-course intensity profiles. ( e ), ( f ): Distribution of IBIs of mouse primary cardiomyocytes. ( e ) The relationship between mean IBIs and fluctuations of beating [coefficient of variability (CV) of IBIs] of single isolated primary cardiomyocytes (blue open circles, n = 73) and primary clusters (red filled triangles, n = 6). ( f ) A histogram of all plots in ( e ). The blue filled bars indicate the frequency of IBIs of single cardiomyocytes; the blue arrow and the error bar indicate the corresponding mean value and standard deviation (SD) of single-cardiomyocyte IBIs, respectively. The red filled bars indicate the frequency of IBIs of clusters; the red arrow and the error bar indicate the corresponding mean value and SD of clusters. ( g ), ( h ): Distribution of IBIs in hES cardiomyocytes. ( g ) The relationship between mean IBIs and CV of IBIs in single isolated hES cardiomyocytes (blue open circles, n = 125) and hES clusters (red filled triangles, n = 27). ( h ) A histogram of all plots in ( g ). The blue filled bars indicate the frequency of IBIs of single cardiomyocytes; the blue arrow and the error bar indicate the corresponding mean values and SD of single cardiomyocytes, respectively. The red filled bars indicate the frequency of IBIs of clusters; the red arrow and the error bar indicate the corresponding mean value and SD of clusters.

Article Snippet: Human embryonic stem cell-derived cardiomyocytes (hES) (hES-CMCTM002, hES cell line SA002) were purchased from Cellectis (Gothenburg, Sweden) , .

Techniques: Isolation, Standard Deviation

Journal: Scientific Reports

Article Title: Emergent synchronous beating behavior in spontaneous beating cardiomyocyte clusters

doi: 10.1038/s41598-021-91466-y

Figure Lengend Snippet: Distribution of IBIs and fluctuation of IBI distribution of the hES cardiomyocyte clusters and their constituent cells. ( a )–( c ): Micrographs of hES cardiomyocyte clusters. ( d )–( f ): Distribution of IBIs and the CV of IBIs in the clusters ( a )–( c ) and isolated constituent cells from each cluster (n=50 from among re-cultivated \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$1.0\times 10^{3}\hbox { cells}$$\end{document} 1.0 × 10 3 cells ). These plots ( d )–( f ) correspond to each cluster ( a )–( c ). The red filled triangles indicate the cardiomyocyte clusters, and the blue open circles indicate constituent cardiomyocytes of each cluster. Each cluster was measured 2 days after the beating started. Single cardiomyocytes were isolated from each cluster by trypsinization. IBIs of single constituent cardiomyocytes were measured 3 days after their isolation. Median and 95% confidence interval of single cardiomyocytes were 0.971 s and 0.825–1.25 s ( d ), 1.19 s and 1.00–1.28 s ( e ), and 1.12 s and 0.844–1.22 s ( f ), respectively. ( g )–( i ): Histograms of IBIs of each cluster and its isolated constituent cells. The blue filled bars indicate the ratio of frequency for single constituent cardiomyocytes; the blue arrows and error bars indicate the mean IBIs and SDs, and the red arrows also indicate the mean IBIs of clusters.

Article Snippet: Human embryonic stem cell-derived cardiomyocytes (hES) (hES-CMCTM002, hES cell line SA002) were purchased from Cellectis (Gothenburg, Sweden) , .

Techniques: Isolation

Journal: Scientific Reports

Article Title: Emergent synchronous beating behavior in spontaneous beating cardiomyocyte clusters

doi: 10.1038/s41598-021-91466-y

Figure Lengend Snippet: Influence of trypsinization on interbeat intervals in dispersed individual hES cardiomyocytes. ( a ) Distribution of the IBIs and the CV of IBIs in single hES cardiomyocytes before and after trypsinization. The blue open circles indicate the single hES cardiomyocytes (n = 50) before trypsinization. The orange open circles indicate the single cardiomyocytes (n = 50) after trypsinization. ( b ) Histograms of IBIs of hES single cardiomyocytes before and after trypsinization. The blue filled bars indicate the mean IBIs of single cardiomyocytes before trypsinization; the blue arrow and error bar indicate their mean value and SD. The orange filled bars indicate the frequency of mean IBIs of trypsinized single cardiomyocytes; the orange arrow and error bar indicate their mean value and SD.

Article Snippet: Human embryonic stem cell-derived cardiomyocytes (hES) (hES-CMCTM002, hES cell line SA002) were purchased from Cellectis (Gothenburg, Sweden) , .

Techniques:

Journal: Scientific Reports

Article Title: Emergent synchronous beating behavior in spontaneous beating cardiomyocyte clusters

doi: 10.1038/s41598-021-91466-y

Figure Lengend Snippet: Distributions of interbeat intervals (IBIs) and fluctuations of the two hES cardiomyocyte clusters before and after their connection and after re-separation. ( a )–( e ): Micrographs of cardiomyocyte clusters. Micrographs of the large cluster ( a ) and small cluster ( b ) before contact. These clusters were measured when they had been cultivated for 7 days. The two hES cardiomyocyte clusters were connected ( c ). The measurement was performed 3 days after the two clusters contacted each other. Micrographs of the large cluster ( d ) and small cluster ( e ) after separation. The measurements were taken within 5 min of separation. ( f ): Distribution of IBIs and fluctuations of two clusters before contact, during contact, and after separation. Blue filled bar and error bar indicate the mean IBIs and SD of the large cluster. Green filled bar and error bar indicate the mean IBIs and SD of the small cluster.

Article Snippet: Human embryonic stem cell-derived cardiomyocytes (hES) (hES-CMCTM002, hES cell line SA002) were purchased from Cellectis (Gothenburg, Sweden) , .

Techniques: