Comparison of cultured human cardiomyocyte clusters obtained from embryos/fetuses or derived from human embryonic stem cells

Søren J Grubb; Maj Linea Vestergaard; Astrid Sten Andersen; Karen Koefod Rasmussen; Linn Mamsen; Greta Tuckute; Kristina Grunnet-Lauridsen; Kjeld Møllgård; Erik Ernst; Søren T Christensen; Kirstine Calloe; Claus Yding Andersen

doi:10.1089/scd.2018.0231

Comparison of cultured human cardiomyocyte clusters obtained from embryos/fetuses or derived from human embryonic stem cells

Søren J Grubb, Maj Linea Vestergaard, Astrid Sten Andersen, Karen Koefod Rasmussen, Linn Mamsen, Greta Tuckute, Kristina Grunnet-Lauridsen, Kjeld Møllgård, Erik Ernst, Søren T Christensen, Kirstine Calloe, Claus Yding Andersen

1 Citation (Scopus)

Abstract

Cardiomyocytes (CMs) derived from human embryonic stem cells (hESCs) or induced pluripotent stem cells (iPSCs) are used to study cardiogenesis and mechanisms of heart disease, and are being used in methods for toxiological screening of drugs. The phenotype of stem-cell-derived CMs should ideally resemble native CMs. Here, we compare embryonic/fetal CMs with hESC-derived CMs according to function and morphology. CM clusters were obtained from human embryonic/fetal hearts from elective terminated pregnancies before gestational week 12, and separated into atrial and ventricular tissues. Specific markers for embryonic CMs and primary cilia were visualized using immunofluorescence microscopy analysis. Contracting human embryonic cardiomyocyte (hECM) clusters morphologically and phenotypically resemble CMs in the embryonic/fetal heart. In addition, the contracting hECM clusters expressed primary cilia similar to that of cells in the embryonic/fetal heart. The electrophysiological characteristics of atrial and ventricular CMs were established by recording action potentials (APs) using sharp electrodes. In contrast to ventricular APs, atrial APs displayed a marked early repolarization followed by a plateau phase. hESC-CMs displayed a continuum of AP shapes. In all embryonic/fetal clusters, both atrial and ventricular, AP duration was prolonged by exposure to the K_V11.1 channel inhibitor dofetilide (50 nM); however, the prolongation was not significant, possibly due to the relatively small number of experiments. This study provides novel information on APs and functional characteristics of atrial and ventricular CMs in first trimester hearts, and demonstrates that K_v11.1 channels play a functional role already at these early stages. These results provide information needed to validate methods being developed on the basis of in vitro-derived CMs from either hESC or iPSC, and although there was a good correlation between the morphology of the two types of CMs, differences in electrophysiological characteristics exist.

Original language	English
Journal	Stem Cells and Development
Volume	28
Issue number	9
Pages (from-to)	608-619
ISSN	1547-3287
DOIs	https://doi.org/10.1089/scd.2018.0231
Publication status	Published - 1 May 2019

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10.1089/scd.2018.0231

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Grubb, S. J., Vestergaard, M. L., Andersen, A. S., Rasmussen, K. K., Mamsen, L., Tuckute, G., Grunnet-Lauridsen, K., Møllgård, K., Ernst, E., Christensen, S. T., Calloe, K., & Andersen, C. Y. (2019). Comparison of cultured human cardiomyocyte clusters obtained from embryos/fetuses or derived from human embryonic stem cells. Stem Cells and Development, 28(9), 608-619. https://doi.org/10.1089/scd.2018.0231

Grubb, SJ, Vestergaard, ML, Andersen, AS, Rasmussen, KK, Mamsen, L, Tuckute, G, Grunnet-Lauridsen, K, Møllgård, K, Ernst, E, Christensen, ST , Calloe, K & Andersen, CY 2019, 'Comparison of cultured human cardiomyocyte clusters obtained from embryos/fetuses or derived from human embryonic stem cells', Stem Cells and Development, vol. 28, no. 9, pp. 608-619. https://doi.org/10.1089/scd.2018.0231

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title = "Comparison of cultured human cardiomyocyte clusters obtained from embryos/fetuses or derived from human embryonic stem cells",

abstract = "Cardiomyocytes (CMs) derived from human embryonic stem cells (hESCs) or induced pluripotent stem cells (iPSCs) are used to study cardiogenesis and mechanisms of heart disease, and are being used in methods for toxiological screening of drugs. The phenotype of stem-cell-derived CMs should ideally resemble native CMs. Here, we compare embryonic/fetal CMs with hESC-derived CMs according to function and morphology. CM clusters were obtained from human embryonic/fetal hearts from elective terminated pregnancies before gestational week 12, and separated into atrial and ventricular tissues. Specific markers for embryonic CMs and primary cilia were visualized using immunofluorescence microscopy analysis. Contracting human embryonic cardiomyocyte (hECM) clusters morphologically and phenotypically resemble CMs in the embryonic/fetal heart. In addition, the contracting hECM clusters expressed primary cilia similar to that of cells in the embryonic/fetal heart. The electrophysiological characteristics of atrial and ventricular CMs were established by recording action potentials (APs) using sharp electrodes. In contrast to ventricular APs, atrial APs displayed a marked early repolarization followed by a plateau phase. hESC-CMs displayed a continuum of AP shapes. In all embryonic/fetal clusters, both atrial and ventricular, AP duration was prolonged by exposure to the KV11.1 channel inhibitor dofetilide (50 nM); however, the prolongation was not significant, possibly due to the relatively small number of experiments. This study provides novel information on APs and functional characteristics of atrial and ventricular CMs in first trimester hearts, and demonstrates that Kv11.1 channels play a functional role already at these early stages. These results provide information needed to validate methods being developed on the basis of in vitro-derived CMs from either hESC or iPSC, and although there was a good correlation between the morphology of the two types of CMs, differences in electrophysiological characteristics exist.",

author = "Grubb, {S{\o}ren J} and Vestergaard, {Maj Linea} and Andersen, {Astrid Sten} and Rasmussen, {Karen Koefod} and Linn Mamsen and Greta Tuckute and Kristina Grunnet-Lauridsen and Kjeld M{\o}llg{\aa}rd and Erik Ernst and Christensen, {S{\o}ren T} and Kirstine Calloe and Andersen, {Claus Yding}",

year = "2019",

month = may,

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doi = "10.1089/scd.2018.0231",

language = "English",

volume = "28",

pages = "608--619",

journal = "Stem Cells and Development",

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TY - JOUR

T1 - Comparison of cultured human cardiomyocyte clusters obtained from embryos/fetuses or derived from human embryonic stem cells

AU - Grubb, Søren J

AU - Vestergaard, Maj Linea

AU - Andersen, Astrid Sten

AU - Rasmussen, Karen Koefod

AU - Mamsen, Linn

AU - Tuckute, Greta

AU - Grunnet-Lauridsen, Kristina

AU - Møllgård, Kjeld

AU - Ernst, Erik

AU - Christensen, Søren T

AU - Calloe, Kirstine

AU - Andersen, Claus Yding

PY - 2019/5/1

Y1 - 2019/5/1

N2 - Cardiomyocytes (CMs) derived from human embryonic stem cells (hESCs) or induced pluripotent stem cells (iPSCs) are used to study cardiogenesis and mechanisms of heart disease, and are being used in methods for toxiological screening of drugs. The phenotype of stem-cell-derived CMs should ideally resemble native CMs. Here, we compare embryonic/fetal CMs with hESC-derived CMs according to function and morphology. CM clusters were obtained from human embryonic/fetal hearts from elective terminated pregnancies before gestational week 12, and separated into atrial and ventricular tissues. Specific markers for embryonic CMs and primary cilia were visualized using immunofluorescence microscopy analysis. Contracting human embryonic cardiomyocyte (hECM) clusters morphologically and phenotypically resemble CMs in the embryonic/fetal heart. In addition, the contracting hECM clusters expressed primary cilia similar to that of cells in the embryonic/fetal heart. The electrophysiological characteristics of atrial and ventricular CMs were established by recording action potentials (APs) using sharp electrodes. In contrast to ventricular APs, atrial APs displayed a marked early repolarization followed by a plateau phase. hESC-CMs displayed a continuum of AP shapes. In all embryonic/fetal clusters, both atrial and ventricular, AP duration was prolonged by exposure to the KV11.1 channel inhibitor dofetilide (50 nM); however, the prolongation was not significant, possibly due to the relatively small number of experiments. This study provides novel information on APs and functional characteristics of atrial and ventricular CMs in first trimester hearts, and demonstrates that Kv11.1 channels play a functional role already at these early stages. These results provide information needed to validate methods being developed on the basis of in vitro-derived CMs from either hESC or iPSC, and although there was a good correlation between the morphology of the two types of CMs, differences in electrophysiological characteristics exist.

AB - Cardiomyocytes (CMs) derived from human embryonic stem cells (hESCs) or induced pluripotent stem cells (iPSCs) are used to study cardiogenesis and mechanisms of heart disease, and are being used in methods for toxiological screening of drugs. The phenotype of stem-cell-derived CMs should ideally resemble native CMs. Here, we compare embryonic/fetal CMs with hESC-derived CMs according to function and morphology. CM clusters were obtained from human embryonic/fetal hearts from elective terminated pregnancies before gestational week 12, and separated into atrial and ventricular tissues. Specific markers for embryonic CMs and primary cilia were visualized using immunofluorescence microscopy analysis. Contracting human embryonic cardiomyocyte (hECM) clusters morphologically and phenotypically resemble CMs in the embryonic/fetal heart. In addition, the contracting hECM clusters expressed primary cilia similar to that of cells in the embryonic/fetal heart. The electrophysiological characteristics of atrial and ventricular CMs were established by recording action potentials (APs) using sharp electrodes. In contrast to ventricular APs, atrial APs displayed a marked early repolarization followed by a plateau phase. hESC-CMs displayed a continuum of AP shapes. In all embryonic/fetal clusters, both atrial and ventricular, AP duration was prolonged by exposure to the KV11.1 channel inhibitor dofetilide (50 nM); however, the prolongation was not significant, possibly due to the relatively small number of experiments. This study provides novel information on APs and functional characteristics of atrial and ventricular CMs in first trimester hearts, and demonstrates that Kv11.1 channels play a functional role already at these early stages. These results provide information needed to validate methods being developed on the basis of in vitro-derived CMs from either hESC or iPSC, and although there was a good correlation between the morphology of the two types of CMs, differences in electrophysiological characteristics exist.

U2 - 10.1089/scd.2018.0231

DO - 10.1089/scd.2018.0231

M3 - Journal article

C2 - 30755084

SN - 1547-3287

VL - 28

SP - 608

EP - 619

JO - Stem Cells and Development

JF - Stem Cells and Development

IS - 9

ER -

Comparison of cultured human cardiomyocyte clusters obtained from embryos/fetuses or derived from human embryonic stem cells

Abstract

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