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Article
Calcium Transient Assays for Compound Screening with Human iPSC-derived Cardiomyocytes: Evaluating New Tools
Journal of Evolving Stem Cell Research
  • Neil J. Daily, InvivoSciences, Inc.
  • Radleigh Santos, Torrey Pines Institute for Molecular Studies
  • Joseph Vecchi, InvivoSciences, Inc.
  • Pinar Kemanli, InvivoSciences, Inc.
  • Tetsuro Wakatsuki, InvivoSciences, Inc.
Document Type
Article
Publication Date
1-24-2017
Keywords
  • Calcium transients,
  • iPSC-derived human cardiomyocytes
Abstract

Calcium (Ca2+) plays a central role in regulating many biological processes in the cell from muscle contraction to neurotransmitter release. The need for reliable fluorescent calcium indicator dyes is of vast importance for studying many aspects of cell biology as well as screening compounds using phenotypic high throughput assays. We have assessed two of the latest generation of calcium indicator dyes, FLIPR Calcium 6 and Cal-520 AM for studying calcium transients (CaTs) in induced pluripotent stem cell (iPSC) -derived human cardiomyocytes. FLIPR Calcium 6 and Cal-520 dyes both displayed robust CaTs with a high signal-to-noise ratio (SNR) and were non-toxic to the cells. The analysis showed that CaT amplitudes were stable between measurements, but CaT duration was more variable and tended to increase between reads. Two methods were compared for drug-screening hit-selection; difference in average (unstandardized) and standardized difference. The unstandardized difference was better for assessing CaT amplitude, whereas standardized difference was equal to or better for assessing CaT duration. In summary, FLIPR Calcium 6 and Cal-520 are suitable dyes for drug-screening using iPSC-derived human cardiomyocytes.

Comments

© 2017 Neil J. Daily, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Additional Comments
NIH grant #s: R43AG052220, R43AG109735, R01HL109505
DOI
10.14302/issn.2574-4372.jesr-16-1395
Citation Information
Neil J. Daily, Radleigh Santos, Joseph Vecchi, Pinar Kemanli, et al.. "Calcium Transient Assays for Compound Screening with Human iPSC-derived Cardiomyocytes: Evaluating New Tools" Journal of Evolving Stem Cell Research Vol. 1 Iss. 2 (2017) p. 1 - 11 ISSN: 2574-4372
Available at: http://works.bepress.com/radleigh-santos/9/