Uninterrupted monitoring of drug effects in human-induced pluripotent stem cell-derived cardiomyocytes with bioluminescence Ca2+ microscopy

BMC Research Notes
Kazushi SuzukiTakeharu Nagai

Abstract

Cardiomyocytes derived from human-induced pluripotent stem cells are a powerful platform for high-throughput drug screening in vitro. However, current modalities for drug testing, such as electrophysiology and fluorescence imaging have inherent drawbacks. To circumvent these problems, we report the development of a bioluminescent Ca2+ indicator GmNL(Ca2+), and its application in a customized microscope for high-throughput drug screening. GmNL(Ca2+) gives a 140% signal change with Ca2+, and can image drug-induced changes of Ca2+ dynamics in cultured cells. Since bioluminescence requires application of a chemical substrate, which is consumed over ~ 30 min we made a dedicated microscope with automated drug dispensing inside a light-tight box, to control drug addition. To overcome thermal instability of the luminescent substrate, or small molecule, dual climate control enables distinct temperature settings in the drug reservoir and the biological sample. By combining GmNL(Ca2+) with this adaptation, we could image spontaneous Ca2+ transients in cultured cardiomyocytes and phenotype their response to well-known drugs without accessing the sample directly. In addition, the bioluminescent strategy demonstrates minimal perturbation of ...Continue Reading

References

Dec 13, 2012·Nature Communications·Kenta SaitoTakeharu Nagai
Feb 2, 2013·Circulation Research·Mark MercolaErik Willems
May 22, 2013·Toxicological Sciences : an Official Journal of the Society of Toxicology·Kate HarrisArun Sridhar
May 9, 2014·Circulation Research·Lars KaestnerPeter Lipp
Apr 2, 2015·Proceedings of the National Academy of Sciences of the United States of America·Akira TakaiTakeharu Nagai
Jul 15, 2015·Journal of Biomolecular Screening·Bernard FerminiJamie I Vandenberg
Dec 15, 2016·Nature Communications·Kazushi SuzukiTakeharu Nagai
Oct 3, 2017·Journal of Evolving Stem Cell Research·Neil J DailyTetsuro Wakatsuki
Jan 2, 2018·Cell Chemical Biology·Hajime ShinodaTakeharu Nagai

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Citations

Mar 17, 2021·ACS Sensors·Brittany R BenlianEvan W Miller

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Methods Mentioned

BETA
fluorescence
fluorescence microscopy
fluorescence imaging
PCR
fluorescence titration assay
FRET
bioluminescence

Software Mentioned

Gamillus
NIS

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