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A non-invasive platform for functional characterization of stem-cell-derived cardiomyocytes with applications in cardiotoxicity testing.

Maddah M, Heidmann JD, Mandegar MA, Walker CD, Bolouki S, Conklin BR, Loewke KE - Stem Cell Reports (2015)

Bottom Line: We present a non-invasive method to characterize the function of pluripotent stem-cell-derived cardiomyocytes based on video microscopy and image analysis.The platform, called Pulse, generates automated measurements of beating frequency, beat duration, amplitude, and beat-to-beat variation based on motion analysis of phase-contrast images captured at a fast frame rate.Using Pulse, we demonstrate recapitulation of drug effects in stem-cell-derived cardiomyocytes without the use of exogenous labels and show that our platform can be used for high-throughput cardiotoxicity drug screening and studying physiologically relevant phenotypes.

View Article: PubMed Central - PubMed

Affiliation: Cellogy, Inc., Palo Alto, CA 94301, USA. Electronic address: mmaddah@alum.mit.edu.

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Drug Testing ExperimentsWe studied and quantified effect of eight drugs by motion analysis of GCaMP cardiomyocytes. The left panel shows the signals estimated from a video of one example region before and after the addition of the drug. The bar charts on the right show the variation of a beating parameter (mean and SEM) for control and different doses at four time points. Each drug concentration included technical replicates of six wells and four recordings per well, for 24 total measurements. The mean values were calculated by first taking the average of within-well replicates, reducing the 24 measurements per concentration from 24 to 6, and then calculating the mean and SEM with n = 6.(A) Norepinephrine. Our method detects a dose-dependent increase in beat rate and decrease in beat duration.(B) Cisapride. We detect arrhythmic beating patterns, an increase in beat duration, and decrease in beat rate.(C) E-4031. We detect an arrhythmic beating patterns, an increase in beat duration, and decrease in beat rate.(D) Verapamil. Our method detects significant decrease in beat duration, as well as decrease in beat rate at high concentrations.(E) Quinidine. Our method detects a dose-dependent increase in beat duration and decrease in beat rate.(F) Sotalol. Our method detects a dose-dependent increase in the beat duration.(G) Nifedipine. Our method detects a dose-dependent increase in the beat duration.(H) Aspirin. We detect no significant change in beat rate, irregularity, or duration.See also Figure S2.
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fig5: Drug Testing ExperimentsWe studied and quantified effect of eight drugs by motion analysis of GCaMP cardiomyocytes. The left panel shows the signals estimated from a video of one example region before and after the addition of the drug. The bar charts on the right show the variation of a beating parameter (mean and SEM) for control and different doses at four time points. Each drug concentration included technical replicates of six wells and four recordings per well, for 24 total measurements. The mean values were calculated by first taking the average of within-well replicates, reducing the 24 measurements per concentration from 24 to 6, and then calculating the mean and SEM with n = 6.(A) Norepinephrine. Our method detects a dose-dependent increase in beat rate and decrease in beat duration.(B) Cisapride. We detect arrhythmic beating patterns, an increase in beat duration, and decrease in beat rate.(C) E-4031. We detect an arrhythmic beating patterns, an increase in beat duration, and decrease in beat rate.(D) Verapamil. Our method detects significant decrease in beat duration, as well as decrease in beat rate at high concentrations.(E) Quinidine. Our method detects a dose-dependent increase in beat duration and decrease in beat rate.(F) Sotalol. Our method detects a dose-dependent increase in the beat duration.(G) Nifedipine. Our method detects a dose-dependent increase in the beat duration.(H) Aspirin. We detect no significant change in beat rate, irregularity, or duration.See also Figure S2.

Mentions: Figure 4A shows the beating signal extracted by Pulse 30 min after addition of different concentrations of blebbistatin. As the concentration of blebbistatin is increased, the relative amplitude of the beating signal decreases, serving as a functional readout for the expected reduction in contractility. Figure 4B shows a comparison of the amplitude of the beating signal before blebbistatin is added, 30 min after compound application, and after a subsequent media change. Note that the biomechanical beating of the cardiomyocytes is restored back to normal levels after blebbistatin is washed away. For a concentration of 1 μM or above, no biomechanical beating is observed. In contrast, fluorescence imaging of GCaMP continues to generate a calcium transient signal even at 5 μM, as seen in Figure 5C. These data demonstrate that Pulse is able to capture and quantify the effect of a myosin II blocker in inhibiting the biomechanical contraction of the cardiomyocytes, an effect that MEA or Ca+2 imaging systems do not capture (Abassi et al., 2012).


A non-invasive platform for functional characterization of stem-cell-derived cardiomyocytes with applications in cardiotoxicity testing.

Maddah M, Heidmann JD, Mandegar MA, Walker CD, Bolouki S, Conklin BR, Loewke KE - Stem Cell Reports (2015)

Drug Testing ExperimentsWe studied and quantified effect of eight drugs by motion analysis of GCaMP cardiomyocytes. The left panel shows the signals estimated from a video of one example region before and after the addition of the drug. The bar charts on the right show the variation of a beating parameter (mean and SEM) for control and different doses at four time points. Each drug concentration included technical replicates of six wells and four recordings per well, for 24 total measurements. The mean values were calculated by first taking the average of within-well replicates, reducing the 24 measurements per concentration from 24 to 6, and then calculating the mean and SEM with n = 6.(A) Norepinephrine. Our method detects a dose-dependent increase in beat rate and decrease in beat duration.(B) Cisapride. We detect arrhythmic beating patterns, an increase in beat duration, and decrease in beat rate.(C) E-4031. We detect an arrhythmic beating patterns, an increase in beat duration, and decrease in beat rate.(D) Verapamil. Our method detects significant decrease in beat duration, as well as decrease in beat rate at high concentrations.(E) Quinidine. Our method detects a dose-dependent increase in beat duration and decrease in beat rate.(F) Sotalol. Our method detects a dose-dependent increase in the beat duration.(G) Nifedipine. Our method detects a dose-dependent increase in the beat duration.(H) Aspirin. We detect no significant change in beat rate, irregularity, or duration.See also Figure S2.
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fig5: Drug Testing ExperimentsWe studied and quantified effect of eight drugs by motion analysis of GCaMP cardiomyocytes. The left panel shows the signals estimated from a video of one example region before and after the addition of the drug. The bar charts on the right show the variation of a beating parameter (mean and SEM) for control and different doses at four time points. Each drug concentration included technical replicates of six wells and four recordings per well, for 24 total measurements. The mean values were calculated by first taking the average of within-well replicates, reducing the 24 measurements per concentration from 24 to 6, and then calculating the mean and SEM with n = 6.(A) Norepinephrine. Our method detects a dose-dependent increase in beat rate and decrease in beat duration.(B) Cisapride. We detect arrhythmic beating patterns, an increase in beat duration, and decrease in beat rate.(C) E-4031. We detect an arrhythmic beating patterns, an increase in beat duration, and decrease in beat rate.(D) Verapamil. Our method detects significant decrease in beat duration, as well as decrease in beat rate at high concentrations.(E) Quinidine. Our method detects a dose-dependent increase in beat duration and decrease in beat rate.(F) Sotalol. Our method detects a dose-dependent increase in the beat duration.(G) Nifedipine. Our method detects a dose-dependent increase in the beat duration.(H) Aspirin. We detect no significant change in beat rate, irregularity, or duration.See also Figure S2.
Mentions: Figure 4A shows the beating signal extracted by Pulse 30 min after addition of different concentrations of blebbistatin. As the concentration of blebbistatin is increased, the relative amplitude of the beating signal decreases, serving as a functional readout for the expected reduction in contractility. Figure 4B shows a comparison of the amplitude of the beating signal before blebbistatin is added, 30 min after compound application, and after a subsequent media change. Note that the biomechanical beating of the cardiomyocytes is restored back to normal levels after blebbistatin is washed away. For a concentration of 1 μM or above, no biomechanical beating is observed. In contrast, fluorescence imaging of GCaMP continues to generate a calcium transient signal even at 5 μM, as seen in Figure 5C. These data demonstrate that Pulse is able to capture and quantify the effect of a myosin II blocker in inhibiting the biomechanical contraction of the cardiomyocytes, an effect that MEA or Ca+2 imaging systems do not capture (Abassi et al., 2012).

Bottom Line: We present a non-invasive method to characterize the function of pluripotent stem-cell-derived cardiomyocytes based on video microscopy and image analysis.The platform, called Pulse, generates automated measurements of beating frequency, beat duration, amplitude, and beat-to-beat variation based on motion analysis of phase-contrast images captured at a fast frame rate.Using Pulse, we demonstrate recapitulation of drug effects in stem-cell-derived cardiomyocytes without the use of exogenous labels and show that our platform can be used for high-throughput cardiotoxicity drug screening and studying physiologically relevant phenotypes.

View Article: PubMed Central - PubMed

Affiliation: Cellogy, Inc., Palo Alto, CA 94301, USA. Electronic address: mmaddah@alum.mit.edu.

Show MeSH
Related in: MedlinePlus