Optical mapping of human embryonic stem cell-derived cardiomyocyte graft electrical activity in injured hearts
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RESEARCH
Open Access
Optical mapping of human embryonic stem cell-derived cardiomyocyte graft electrical activity in injured hearts Dominic Filice1,2†, Wahiba Dhahri3,4†, Joell L. Solan5, Paul D. Lampe5, Erin Steele2,6, Nikita Milani1,2, Benjamin Van Biber2,7, Wei-Zhong Zhu2,7, Tamilla Sadikov Valdman3,4, Rocco Romagnolo3,4, José David Otero-Cruz2,7, Kip D. Hauch1, Matthew W. Kay8, Narine Sarvazyan9 and Michael A. Laflamme3,4,10*
Abstract Background: Human embryonic stem cell-derived cardiomyocytes (hESC-CMs) show tremendous promise for cardiac regeneration, but the successful development of hESC-CM-based therapies requires improved tools to investigate their electrical behavior in recipient hearts. While optical voltage mapping is a powerful technique for studying myocardial electrical activity ex vivo, we have previously shown that intra-cardiac hESC-CM grafts are not labeled by conventional voltage-sensitive fluorescent dyes. We hypothesized that the water-soluble voltagesensitive dye di-2-ANEPEQ would label engrafted hESC-CMs and thereby facilitate characterization of graft electrical function and integration. Methods: We developed and validated a novel optical voltage mapping strategy based on the simultaneous imaging of the calcium-sensitive fluorescent protein GCaMP3, a graft-autonomous reporter of graft activation, and optical action potentials (oAPs) derived from di-2-ANEPEQ, which labels both graft and host myocardium. Cardiomyocytes from three different GCaMP3+ hESC lines (H7, RUES2, or ESI-17) were transplanted into guinea pig models of subacute and chronic infarction, followed by optical mapping at 2 weeks post-transplantation. Results: Use of a water-soluble voltage-sensitive dye revealed pro-arrhythmic properties of GCaMP3+ hESC-CM grafts from all three lines including slow conduction velocity, incomplete host-graft coupling, and spatially heterogeneous patterns of activation that varied beat-to-beat. GCaMP3+ hESC-CMs from the RUES2 and ESI-17 lines both showed prolonged oAP durations both in vitro and in vivo. Although hESC-CMs partially remuscularize the injured hearts, histological evaluation revealed immature graft structure and impaired gap junction expression at this early timepoint. (Continued on next page)
* Correspondence: [email protected] † Dominic Filice and Wahiba Dhahri contributed equally to this work. 3 McEwen Stem Cell Institute, University Health Network, 101 College Street, Rm 3-908, Toronto, ON M5G 1L7, Canada 4 Peter Munk Cardiac Centre, University Health Network, Toronto, ON M5G 2N2, Canada Full list of author information is available at the end of the article © The Author(s). 2020 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third
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