Engraftment of human embryonic stem cell derived cardiomyocytes improves conduction in an arrhythmogenic in vitro model

Susan A. Thompson, Paul W. Burridge, Elizabeth A. Lipke, Michael Shamblott, Elias Zambidis, Leslie Tung

Research output: Contribution to journalArticle

Abstract

In this study, we characterized the electrophysiological benefits of engrafting human embryonic stem cell-derived cardiomyocytes (hESC-CMs) in a model of arrhythmogenic cardiac tissue. Using transforming growth factor-β treated monolayers of neonatal rat ventricular cells (NRVCs), which retain several key aspects of the healing infarct such as an excess of contractile myofibroblasts and slowed, heterogeneous conduction, we assessed the ability of hESC-CMs to improve conduction and prevent arrhythmias. Cells from beating embryoid bodies (hESC-CMs) can form functional monolayers which beat spontaneously and can be electrically stimulated, with mean action potential duration of 275 ± 36. ms and conduction velocity (CV) of 10.6 ± 4.2. cm/s (n = 3). These cells, or cells from non-beating embryoid bodies (hEBCs) were added to anisotropic, NRVC monolayers. Immunostaining demonstrated hESC-CM survival and engraftment, and dye transfer assays confirmed functional coupling between hESC-CMs and NRVCs. Conduction velocities significantly increased in anisotropic NRVC monolayers after engraftment of hESC-CMs (13.4 ± 0.9. cm/s, n = 35 vs. 30.1 ± 3.2. cm/s, n = 20 in the longitudinal direction and 4.3 ± 0.3. cm/s vs. 9.3 ± 0.9. cm/s in the transverse direction), but decreased to even lower values after engraftment of non-cardiac hEBCs (to 10.6 ± 1.3. cm/s and 3.1 ± 0.5. cm/s, n = 11, respectively). Furthermore, reentrant wave vulnerability in NRVC monolayers decreased by 20% after engraftment of hESC-CMs, but did not change with engraftment of hEBCs. Finally, the culture of hESC-CMs in transwell inserts, which prevents juxtacrine interactions, or engraftment with connexin43-silenced hESC-CMs provided no functional improvement to NRVC monolayers. These results demonstrate that hESC-CMs can reverse the slowing of conduction velocity, reduce the incidence of reentry, and augment impaired electrical propagation via gap junction coupling to host cardiomyocytes in this arrhythmogenic in vitro model.

Original languageEnglish (US)
Pages (from-to)15-23
Number of pages9
JournalJournal of Molecular and Cellular Cardiology
Volume53
Issue number1
DOIs
StatePublished - Jul 2012

Fingerprint

Cardiac Myocytes
Embryoid Bodies
In Vitro Techniques
Human Embryonic Stem Cells
Connexin 43
Myofibroblasts
Gap Junctions
Transforming Growth Factors
Action Potentials
Cardiac Arrhythmias
Coloring Agents
Incidence

Keywords

  • Arrhythmia
  • Cardiomyocytes
  • Electrophysiology
  • Human embryonic stem cell
  • Myocardial infarction

ASJC Scopus subject areas

  • Molecular Biology
  • Cardiology and Cardiovascular Medicine

Cite this

Engraftment of human embryonic stem cell derived cardiomyocytes improves conduction in an arrhythmogenic in vitro model. / Thompson, Susan A.; Burridge, Paul W.; Lipke, Elizabeth A.; Shamblott, Michael; Zambidis, Elias; Tung, Leslie.

In: Journal of Molecular and Cellular Cardiology, Vol. 53, No. 1, 07.2012, p. 15-23.

Research output: Contribution to journalArticle

@article{3a6beda8b0f445b39fd007e6b380bddc,
title = "Engraftment of human embryonic stem cell derived cardiomyocytes improves conduction in an arrhythmogenic in vitro model",
abstract = "In this study, we characterized the electrophysiological benefits of engrafting human embryonic stem cell-derived cardiomyocytes (hESC-CMs) in a model of arrhythmogenic cardiac tissue. Using transforming growth factor-β treated monolayers of neonatal rat ventricular cells (NRVCs), which retain several key aspects of the healing infarct such as an excess of contractile myofibroblasts and slowed, heterogeneous conduction, we assessed the ability of hESC-CMs to improve conduction and prevent arrhythmias. Cells from beating embryoid bodies (hESC-CMs) can form functional monolayers which beat spontaneously and can be electrically stimulated, with mean action potential duration of 275 ± 36. ms and conduction velocity (CV) of 10.6 ± 4.2. cm/s (n = 3). These cells, or cells from non-beating embryoid bodies (hEBCs) were added to anisotropic, NRVC monolayers. Immunostaining demonstrated hESC-CM survival and engraftment, and dye transfer assays confirmed functional coupling between hESC-CMs and NRVCs. Conduction velocities significantly increased in anisotropic NRVC monolayers after engraftment of hESC-CMs (13.4 ± 0.9. cm/s, n = 35 vs. 30.1 ± 3.2. cm/s, n = 20 in the longitudinal direction and 4.3 ± 0.3. cm/s vs. 9.3 ± 0.9. cm/s in the transverse direction), but decreased to even lower values after engraftment of non-cardiac hEBCs (to 10.6 ± 1.3. cm/s and 3.1 ± 0.5. cm/s, n = 11, respectively). Furthermore, reentrant wave vulnerability in NRVC monolayers decreased by 20{\%} after engraftment of hESC-CMs, but did not change with engraftment of hEBCs. Finally, the culture of hESC-CMs in transwell inserts, which prevents juxtacrine interactions, or engraftment with connexin43-silenced hESC-CMs provided no functional improvement to NRVC monolayers. These results demonstrate that hESC-CMs can reverse the slowing of conduction velocity, reduce the incidence of reentry, and augment impaired electrical propagation via gap junction coupling to host cardiomyocytes in this arrhythmogenic in vitro model.",
keywords = "Arrhythmia, Cardiomyocytes, Electrophysiology, Human embryonic stem cell, Myocardial infarction",
author = "Thompson, {Susan A.} and Burridge, {Paul W.} and Lipke, {Elizabeth A.} and Michael Shamblott and Elias Zambidis and Leslie Tung",
year = "2012",
month = "7",
doi = "10.1016/j.yjmcc.2012.01.023",
language = "English (US)",
volume = "53",
pages = "15--23",
journal = "Journal of Molecular and Cellular Cardiology",
issn = "0022-2828",
publisher = "Academic Press Inc.",
number = "1",

}

TY - JOUR

T1 - Engraftment of human embryonic stem cell derived cardiomyocytes improves conduction in an arrhythmogenic in vitro model

AU - Thompson, Susan A.

AU - Burridge, Paul W.

AU - Lipke, Elizabeth A.

AU - Shamblott, Michael

AU - Zambidis, Elias

AU - Tung, Leslie

PY - 2012/7

Y1 - 2012/7

N2 - In this study, we characterized the electrophysiological benefits of engrafting human embryonic stem cell-derived cardiomyocytes (hESC-CMs) in a model of arrhythmogenic cardiac tissue. Using transforming growth factor-β treated monolayers of neonatal rat ventricular cells (NRVCs), which retain several key aspects of the healing infarct such as an excess of contractile myofibroblasts and slowed, heterogeneous conduction, we assessed the ability of hESC-CMs to improve conduction and prevent arrhythmias. Cells from beating embryoid bodies (hESC-CMs) can form functional monolayers which beat spontaneously and can be electrically stimulated, with mean action potential duration of 275 ± 36. ms and conduction velocity (CV) of 10.6 ± 4.2. cm/s (n = 3). These cells, or cells from non-beating embryoid bodies (hEBCs) were added to anisotropic, NRVC monolayers. Immunostaining demonstrated hESC-CM survival and engraftment, and dye transfer assays confirmed functional coupling between hESC-CMs and NRVCs. Conduction velocities significantly increased in anisotropic NRVC monolayers after engraftment of hESC-CMs (13.4 ± 0.9. cm/s, n = 35 vs. 30.1 ± 3.2. cm/s, n = 20 in the longitudinal direction and 4.3 ± 0.3. cm/s vs. 9.3 ± 0.9. cm/s in the transverse direction), but decreased to even lower values after engraftment of non-cardiac hEBCs (to 10.6 ± 1.3. cm/s and 3.1 ± 0.5. cm/s, n = 11, respectively). Furthermore, reentrant wave vulnerability in NRVC monolayers decreased by 20% after engraftment of hESC-CMs, but did not change with engraftment of hEBCs. Finally, the culture of hESC-CMs in transwell inserts, which prevents juxtacrine interactions, or engraftment with connexin43-silenced hESC-CMs provided no functional improvement to NRVC monolayers. These results demonstrate that hESC-CMs can reverse the slowing of conduction velocity, reduce the incidence of reentry, and augment impaired electrical propagation via gap junction coupling to host cardiomyocytes in this arrhythmogenic in vitro model.

AB - In this study, we characterized the electrophysiological benefits of engrafting human embryonic stem cell-derived cardiomyocytes (hESC-CMs) in a model of arrhythmogenic cardiac tissue. Using transforming growth factor-β treated monolayers of neonatal rat ventricular cells (NRVCs), which retain several key aspects of the healing infarct such as an excess of contractile myofibroblasts and slowed, heterogeneous conduction, we assessed the ability of hESC-CMs to improve conduction and prevent arrhythmias. Cells from beating embryoid bodies (hESC-CMs) can form functional monolayers which beat spontaneously and can be electrically stimulated, with mean action potential duration of 275 ± 36. ms and conduction velocity (CV) of 10.6 ± 4.2. cm/s (n = 3). These cells, or cells from non-beating embryoid bodies (hEBCs) were added to anisotropic, NRVC monolayers. Immunostaining demonstrated hESC-CM survival and engraftment, and dye transfer assays confirmed functional coupling between hESC-CMs and NRVCs. Conduction velocities significantly increased in anisotropic NRVC monolayers after engraftment of hESC-CMs (13.4 ± 0.9. cm/s, n = 35 vs. 30.1 ± 3.2. cm/s, n = 20 in the longitudinal direction and 4.3 ± 0.3. cm/s vs. 9.3 ± 0.9. cm/s in the transverse direction), but decreased to even lower values after engraftment of non-cardiac hEBCs (to 10.6 ± 1.3. cm/s and 3.1 ± 0.5. cm/s, n = 11, respectively). Furthermore, reentrant wave vulnerability in NRVC monolayers decreased by 20% after engraftment of hESC-CMs, but did not change with engraftment of hEBCs. Finally, the culture of hESC-CMs in transwell inserts, which prevents juxtacrine interactions, or engraftment with connexin43-silenced hESC-CMs provided no functional improvement to NRVC monolayers. These results demonstrate that hESC-CMs can reverse the slowing of conduction velocity, reduce the incidence of reentry, and augment impaired electrical propagation via gap junction coupling to host cardiomyocytes in this arrhythmogenic in vitro model.

KW - Arrhythmia

KW - Cardiomyocytes

KW - Electrophysiology

KW - Human embryonic stem cell

KW - Myocardial infarction

UR - http://www.scopus.com/inward/record.url?scp=84861971040&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84861971040&partnerID=8YFLogxK

U2 - 10.1016/j.yjmcc.2012.01.023

DO - 10.1016/j.yjmcc.2012.01.023

M3 - Article

C2 - 22713758

AN - SCOPUS:84861971040

VL - 53

SP - 15

EP - 23

JO - Journal of Molecular and Cellular Cardiology

JF - Journal of Molecular and Cellular Cardiology

SN - 0022-2828

IS - 1

ER -