Optical mapping of human embryonic stem cell-derived cardiomyocyte graft electrical activity in injured hearts

Stem Cell Research & Therapy

Kip D. Hauch¹  Dominic Filice²  Nikita Milani²  Matthew W. Kay³  Narine Sarvazyan⁴  Paul D. Lampe⁵  Joell L. Solan⁵  Erin Steele⁶  José David Otero-Cruz⁷  Wei-Zhong Zhu⁷  Benjamin Van Biber⁷  Tamilla Sadikov Valdman⁸  Rocco Romagnolo⁸  Wahiba Dhahri⁸  Michael A. Laflamme⁹ 

[1] Department of Bioengineering, University of Washington, 98195, Seattle, WA, USA;Department of Bioengineering, University of Washington, 98195, Seattle, WA, USA;Institute for Stem Cell & Regenerative Medicine, University of Washington, 98195, Seattle, WA, USA;Department of Biomedical Engineering, G. Washington University, 20052, Washington, DC, USA;Department of Pharmacology & Physiology, G. Washington University, 20052, Washington, DC, USA;Fred Hutchinson Cancer Research Center, 98109, Seattle, WA, USA;Institute for Stem Cell & Regenerative Medicine, University of Washington, 98195, Seattle, WA, USA;Department of Biology, University of Washington, 98195, Seattle, WA, USA;Institute for Stem Cell & Regenerative Medicine, University of Washington, 98195, Seattle, WA, USA;Department of Pathology, University of Washington, 98195, Seattle, WA, USA;McEwen Stem Cell Institute, University Health Network, 101 College Street, Rm 3-908, M5G 1L7, Toronto, ON, Canada;Peter Munk Cardiac Centre, University Health Network, M5G 2N2, Toronto, ON, Canada;McEwen Stem Cell Institute, University Health Network, 101 College Street, Rm 3-908, M5G 1L7, Toronto, ON, Canada;Peter Munk Cardiac Centre, University Health Network, M5G 2N2, Toronto, ON, Canada;Department of Laboratory Medicine & Pathobiology, University of Toronto, M5G 1L7, Toronto, ON, Canada;

关键词: Human embryonic stem cells;    Cardiomyocyte;    Cell transplantation;    Optical mapping;    Cardiac electrophysiology;   

DOI  :  10.1186/s13287-020-01919-w

来源: Springer

【 摘 要 】

BackgroundHuman 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 voltage-sensitive dye di-2-ANEPEQ would label engrafted hESC-CMs and thereby facilitate characterization of graft electrical function and integration.MethodsWe 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.ResultsUse 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.ConclusionSimultaneous imaging of GCaMP3 and di-2-ANEPEQ allowed us to acquire the first unambiguously graft-derived oAPs from hESC-CM-engrafted hearts and yielded critical insights into their arrhythmogenic potential and line-to-line variation.

【 授权许可】

CC BY