Journal: Stem Cells International

Article Title: Optical Recording of Action Potentials in Human Induced Pluripotent Stem Cell-Derived Cardiac Single Cells and Monolayers Generated from Long QT Syndrome Type 1 Patients

doi: 10.1155/2019/7532657

Figure Lengend Snippet: Patch clamp analysis of cardiomyocytes from control- and LQT-iPSC lines. (a) Representative APs of 1 Hz paced a control-iPSC-CM and an LQT-iPSC-CM from II-2 in . Ten consecutive waves are shown. (b) MDP, APA, and APD 90 from cardiomyocytes derived from the six lines: 201B7 ( n = 6), 409B2 ( n = 5), 692D2 ( n = 6), LQT1A1 ( n = 5), LQT1B1 ( n = 5), and LQT1C1 ( n = 7). Data are represented as means ± SEM; ∗∗ p < 0.005. (c) Representative current traces from control- and LQT1-iPSC-CMs. Upper, the protocol in current clamp recording. Middle, representative traces before and after perfusion with 3R4S-chromanol 293B (30 μ mol/l). Lower, 3R4S-chromanol 293B-subtraction. (d) I-V plots of I Ks at the end of the depolarizing step. 692D2 (control) ( n = 3), LQT1B1 ( n = 3); ∗ p < 0.05.

Article Snippet: Binning was set to 4 × 4, and the measurement interval was every 4 ms. Cardiomyocyte monolayers 7–10 days after seeding on each well were stimulated at 1 Hz with 1 ms depolarizing pulses at 10 V. Analysis of APD 90 was performed with FDSS Waveform Analysis software for cardiomyocytes (U8524-12; Hamamatsu Photonics).

Techniques: Patch Clamp, Control, Derivative Assay

Journal: Stem Cells International

Article Title: Optical Recording of Action Potentials in Human Induced Pluripotent Stem Cell-Derived Cardiac Single Cells and Monolayers Generated from Long QT Syndrome Type 1 Patients

doi: 10.1155/2019/7532657

Figure Lengend Snippet: AP measurements of single cells using a membrane potential dye. (a) Examples of AP waves from control (409B2) measured by the dye. ROIs (1)–(5) are ventricular type. ROI (5) (green wave) shows long APD. (b) Examples of AP waves from LQT1B1. ROIs (1)–(4) are ventricular type with long APD. ROI (4) (purple wave) shows EADs. ROI (5) (green wave) is nodal type. (c) Proportion of subtypes in 3 control- and 3 LQT1-iPSC-CMs: 201B7 ( n = 128), 409B2 ( n = 41), 692D2 ( n = 92), LQT1A1 ( n = 192), LQT1B1 ( n = 101), and LQT1C1 ( n = 202). Ventricular-type cells predominate in all six lines. (d) Representative EADs in a ventricular-type cell from LQT1B1. The EADs were defined as over 20% of the AP amplitude. Arrows show EADs. (e) Occurrence ratio of EADs in ventricular cardiomyocytes from the six lines. The number of ventricular cells was used as the denominator; ∗ p < 0.05.

Article Snippet: Binning was set to 4 × 4, and the measurement interval was every 4 ms. Cardiomyocyte monolayers 7–10 days after seeding on each well were stimulated at 1 Hz with 1 ms depolarizing pulses at 10 V. Analysis of APD 90 was performed with FDSS Waveform Analysis software for cardiomyocytes (U8524-12; Hamamatsu Photonics).

Techniques: Membrane, Control

Journal: Stem Cells International

Article Title: Optical Recording of Action Potentials in Human Induced Pluripotent Stem Cell-Derived Cardiac Single Cells and Monolayers Generated from Long QT Syndrome Type 1 Patients

doi: 10.1155/2019/7532657

Figure Lengend Snippet: AP measurements in a cardiomyocyte monolayer. Data are represented as means ± SEM. (a) An example phase contrast image of the monolayer culture. Scale bar, 500 μ m. (b) Representative AP of a paced control-iPSC-CM monolayer. APD 90 was calculated from the average of 10 consecutive waves. (c) APD 90 from 3 control- and 3 LQT-iPSC-CM monolayers: 201B7 ( n = 3), 409B2 ( n = 3), 692D2 ( n = 3), LQT1A1 ( n = 3), LQT1B1 ( n = 3), and LQT1C1 ( n = 3). LQT-iPSC-CMs showed longer APD than controls; ∗∗ p < 0.005.

Article Snippet: Binning was set to 4 × 4, and the measurement interval was every 4 ms. Cardiomyocyte monolayers 7–10 days after seeding on each well were stimulated at 1 Hz with 1 ms depolarizing pulses at 10 V. Analysis of APD 90 was performed with FDSS Waveform Analysis software for cardiomyocytes (U8524-12; Hamamatsu Photonics).

Techniques: Control

Journal: Stem Cells International

Article Title: Optical Recording of Action Potentials in Human Induced Pluripotent Stem Cell-Derived Cardiac Single Cells and Monolayers Generated from Long QT Syndrome Type 1 Patients

doi: 10.1155/2019/7532657

Figure Lengend Snippet: AP measurements in cardiomyocyte monolayers using a high-throughput plate reader. (a) Representative AP waves on a 96-well plate. Pacing at 1 Hz starts 10 s after recording. (b) Cisapride prolonged APD 90 of both control- and LQT1-iPSC-CMs; 692D2 (control) ( n = 3), LQT1B1 ( n = 5). (c) Cisapride prolonged APD 90 of both control- and LQT1-iPSC-CMs with 100 nM isoproterenol; 692D2 ( n = 5), LQT1B1 ( n = 3). (d) Erythromycin prolonged APD 90 of both control- and LQT1-iPSC-CMs with 100 nM isoproterenol; 692D2 ( n = 4), LQT1B1 ( n = 3). LQT-iPSC-CM monolayers in 300 nM and 1000 nM cisapride were not synchronized at 1 Hz pacing. Data are represented as means ± SEM; ∗∗ p < 0.005, ∗ p < 0.05.

Article Snippet: Binning was set to 4 × 4, and the measurement interval was every 4 ms. Cardiomyocyte monolayers 7–10 days after seeding on each well were stimulated at 1 Hz with 1 ms depolarizing pulses at 10 V. Analysis of APD 90 was performed with FDSS Waveform Analysis software for cardiomyocytes (U8524-12; Hamamatsu Photonics).

Techniques: High Throughput Screening Assay, Control