TY - JOUR
T1 - Molecular and electrophysiological evaluation of human cardiomyocyte subtypes to facilitate generation of composite cardiac models
AU - Li, Jiuru
AU - Wiesinger, Alexandra
AU - Fokkert, Lianne
AU - Boukens, Bastiaan J.
AU - Verkerk, Arie O.
AU - Christoffels, Vincent M.
AU - Boink, Gerard J. J.
AU - Devalla, Harsha D.
N1 - Funding Information: The author(s) disclosed receipt of the following financial support for the research, authorship, and/or publication of this article: This work is supported by funding from European Research council starting grant 714866 and associated proof-of-concept grant 899422, Health Holland LentiPace II, Horizon 2020 Eurostars (E114245 and E115484), Dutch Research Council Open Technology Program 18485 to GJJB; Netherlands Organization for Health Research and Development (ZonMW), ZonMw TOP 40-00812-98-17061 to VMC, ZonMw and the Dutch Heart foundation MKMD grant 114021512 and Dutch Heart Foundation Dekker fellowship 2020T023 to HDD. Publisher Copyright: © The Author(s) 2022.
PY - 2022
Y1 - 2022
N2 - Paucity of physiologically relevant cardiac models has limited the widespread application of human induced pluripotent stem cell (hiPSC)-derived cardiomyocytes in drug development. Here, we performed comprehensive characterization of hiPSC-derived cardiomyocyte subtypes from 2D and 3D cultures and established a novel 3D model to study impulse initiation and propagation. Directed differentiation approaches were used to generate sinoatrial nodal (SANCM), atrial (ACM) and ventricular cardiomyocytes (VCM). Single cell RNA sequencing established that the protocols yield distinct cell populations in line with expected identities, which was also confirmed by electrophysiological characterization. In 3D EHT cultures of all subtypes, we observed prominent expression of stretch-responsive genes such as NPPA. Response to rate modulating drugs noradrenaline, carbachol and ivabradine were comparable in single cells and EHTs. Differences in the speed of impulse propagation between the subtypes were more pronounced in EHTs compared with 2D monolayers owing to a progressive increase in conduction velocities in atrial and ventricular cardiomyocytes, in line with a more mature phenotype. In a novel binary EHT model of pacemaker-atrial interface, the SANCM end of the tissue consistently paced the EHTs under baseline conditions, which was inhibited by ivabradine. Taken together, our data provide comprehensive insights into molecular and electrophysiological properties of hiPSC-derived cardiomyocyte subtypes, facilitating the creation of next generation composite cardiac models for drug discovery, disease modeling and cell-based regenerative therapies.
AB - Paucity of physiologically relevant cardiac models has limited the widespread application of human induced pluripotent stem cell (hiPSC)-derived cardiomyocytes in drug development. Here, we performed comprehensive characterization of hiPSC-derived cardiomyocyte subtypes from 2D and 3D cultures and established a novel 3D model to study impulse initiation and propagation. Directed differentiation approaches were used to generate sinoatrial nodal (SANCM), atrial (ACM) and ventricular cardiomyocytes (VCM). Single cell RNA sequencing established that the protocols yield distinct cell populations in line with expected identities, which was also confirmed by electrophysiological characterization. In 3D EHT cultures of all subtypes, we observed prominent expression of stretch-responsive genes such as NPPA. Response to rate modulating drugs noradrenaline, carbachol and ivabradine were comparable in single cells and EHTs. Differences in the speed of impulse propagation between the subtypes were more pronounced in EHTs compared with 2D monolayers owing to a progressive increase in conduction velocities in atrial and ventricular cardiomyocytes, in line with a more mature phenotype. In a novel binary EHT model of pacemaker-atrial interface, the SANCM end of the tissue consistently paced the EHTs under baseline conditions, which was inhibited by ivabradine. Taken together, our data provide comprehensive insights into molecular and electrophysiological properties of hiPSC-derived cardiomyocyte subtypes, facilitating the creation of next generation composite cardiac models for drug discovery, disease modeling and cell-based regenerative therapies.
KW - cardiac differentiation
KW - electrophysiology
KW - engineered heart tissues
KW - iPSC
KW - stem cell
UR - http://www.scopus.com/inward/record.url?scp=85140224396&partnerID=8YFLogxK
U2 - https://doi.org/10.1177/20417314221127908
DO - https://doi.org/10.1177/20417314221127908
M3 - Article
C2 - 36277058
SN - 2041-7314
VL - 13
JO - Journal of Tissue Engineering
JF - Journal of Tissue Engineering
ER -