Optogenetic control of transiently transfected hiPSC-derived cardiomyocytes for the assessment of drug related cardiotoxicity
-
1
Ncardia (Netherlands), Netherlands
-
2
Ncardia (Germany), Germany
Human induced pluripotent stem cell-derived cardiomyocytes (hiPSCMs) are a well-suited and widely accepted model to study drug efficacy and induced cardiac side effects. As electrophysiological responses and the effects of drugs on cardiac ion channels can be strongly beat rate-dependent, the possibility of pacing cells to physiologically and pathophysiologically relevant frequencies is essential. However, one major drawback of electrical stimulation using MEA technology is the generation of stimulation artefacts, impeding the simultaneous recording of electrical signals derived from the cells. Optogenetic functionalization of excitable cells provides an attractive alternative and can be used for investigation of frequency-dependent responses of e.g. anti-arrhythmic drugs on cardiomyocytes. To enable optogenetic functionalization of hiPSCMs, we have developed a technology (the Xpress.4U™ LightPace kit) which achieves transient (over-)expression of the light-gated ion channel Channelrhodopsin-2 (ChR2) in hiPSCMs .
Using the Xpress.4UTM LightPace kit, ChR2 was expressed in Cor.4U® hiPSCMs that were cultured on an Axion Maestro MEA LUMOS plate. The cells were ready for optical stimulation with the Axion Maestro MEA in combination with the LUMOS light delivery device 24 hours post transfection. Optimal optical excitability was observed 2 days post transfection. High frequency fidelity could be achieved in all transfected wells, with cells following stimulation frequencies up to 3 Hz when applying light pulses of 10 ms and 50% light intensity (approx. 1.9 mW/mm2). The cells also exhibited steadily shortening field potential durations with increasing beat rate, ranging from 230±13 ms (for 1.5 Hz, avg.±s.d., n = 22), over 215±13 ms (for 2 Hz) and 193±7 ms (for 2.5 Hz), down to 174±9 ms (for 3 Hz). These findings demonstrate, that the Xpress.4U™ LightPace kit in combination with Axion MEA LUMOS platform provides a suitable tool for the accurate control of cardiomyocyte contraction and therefore, for the assessment of frequency-related drug effects.
Keywords:
hiPSC-derived cardiomyocytes,
Microelectrode Array (MEA),
optogenetics,
channelrhodopsin,
pacing
Conference:
MEA Meeting 2018 | 11th International Meeting on Substrate Integrated Microelectrode Arrays, Reutlingen, Germany, 4 Jul - 6 Jul, 2018.
Presentation Type:
Poster Presentation
Topic:
Stem cell-derived applications
Citation:
Wolters
B,
Nacken
P and
Kettenhofen
RF
(2019). Optogenetic control of transiently transfected hiPSC-derived cardiomyocytes for the assessment of drug related cardiotoxicity.
Conference Abstract:
MEA Meeting 2018 | 11th International Meeting on Substrate Integrated Microelectrode Arrays.
doi: 10.3389/conf.fncel.2018.38.00003
Copyright:
The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers.
They are made available through the Frontiers publishing platform as a service to conference organizers and presenters.
The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated.
Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed.
For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions.
Received:
16 Mar 2018;
Published Online:
17 Jan 2019.
*
Correspondence:
Dr. Benjamin Wolters, Ncardia (Netherlands), Leiden, Netherlands, benjamin.wolters@ncardia.com