Limits...
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.

Show MeSH

Related in: MedlinePlus

Overlay of Beating Signals Derived from Pulse and Patch ClampPatch-clamping data and video were collected concurrently. Motion-analysis-derived and electrophysiologically generated traces for three independent cell cultures (A–C) with varying frequency and irregularity are overlaid, showing similar results.
© Copyright Policy - CC BY-NC-ND
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4400609&req=5

fig3: Overlay of Beating Signals Derived from Pulse and Patch ClampPatch-clamping data and video were collected concurrently. Motion-analysis-derived and electrophysiologically generated traces for three independent cell cultures (A–C) with varying frequency and irregularity are overlaid, showing similar results.

Mentions: We designed a series of experiments to validate the Pulse platform and demonstrate its ability to detect and measure drug-induced changes to cardiomyocyte function. In a first experiment, we compared the results of Pulse’s motion analysis to data collected by simultaneous manual patch clamp (ChanTest Corporation; see experiment protocol in the Supplemental Experimental Procedures). Phase-contrast imaging and patch-clamp recordings were performed simultaneously on iPSC-derived cardiomyocytes (Cor.4U cell line by Axiogenesis). Figure 3 shows the overlay of the signals measured by these two approaches on three independent cardiomyocyte samples. As observed in the figure, the Pulse beating profiles precisely match the corresponding patch-clamp signals in terms of beating patterns and beat duration. These results confirm that the Pulse system is adequately sampling the beating motion and is generating a reliable output. The manual patch-clamp data were collected while simultaneously capturing a video sequence with a known sampling rate. However, we were not able to record absolute time stamps for both data sets, so the resulting traces were manually aligned.


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)

Overlay of Beating Signals Derived from Pulse and Patch ClampPatch-clamping data and video were collected concurrently. Motion-analysis-derived and electrophysiologically generated traces for three independent cell cultures (A–C) with varying frequency and irregularity are overlaid, showing similar results.
© Copyright Policy - CC BY-NC-ND
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4400609&req=5

fig3: Overlay of Beating Signals Derived from Pulse and Patch ClampPatch-clamping data and video were collected concurrently. Motion-analysis-derived and electrophysiologically generated traces for three independent cell cultures (A–C) with varying frequency and irregularity are overlaid, showing similar results.
Mentions: We designed a series of experiments to validate the Pulse platform and demonstrate its ability to detect and measure drug-induced changes to cardiomyocyte function. In a first experiment, we compared the results of Pulse’s motion analysis to data collected by simultaneous manual patch clamp (ChanTest Corporation; see experiment protocol in the Supplemental Experimental Procedures). Phase-contrast imaging and patch-clamp recordings were performed simultaneously on iPSC-derived cardiomyocytes (Cor.4U cell line by Axiogenesis). Figure 3 shows the overlay of the signals measured by these two approaches on three independent cardiomyocyte samples. As observed in the figure, the Pulse beating profiles precisely match the corresponding patch-clamp signals in terms of beating patterns and beat duration. These results confirm that the Pulse system is adequately sampling the beating motion and is generating a reliable output. The manual patch-clamp data were collected while simultaneously capturing a video sequence with a known sampling rate. However, we were not able to record absolute time stamps for both data sets, so the resulting traces were manually aligned.

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