Action potential variability in human pluripotent stem cell-derived cardiomyocytes obtained from healthy donors

A B Carvalho; Keyla Cristiny da Silva Coutinho; Raiana Andrade Quintanilha Barbosa; Dilza Balteiro Pereira de Campos; Isabela de Carvalho Leitão; R S Pinto; D Silva Dos Santos; Bruna Farjun; Dayana da Silva De Araújo; Fernanda Cristina Paccola Mesquita; G Monnerat-Cahli; E H Medei; Tais Hanae Kasai-Brunswick; A C Campos De Carvalho

doi:10.3389/fphys.2022.1077069

Action potential variability in human pluripotent stem cell-derived cardiomyocytes obtained from healthy donors

Front Physiol. 2022 Dec 16:13:1077069. doi: 10.3389/fphys.2022.1077069. eCollection 2022.

Authors

A B Carvalho^{1

2

3}, Keyla Cristiny da Silva Coutinho¹, Raiana Andrade Quintanilha Barbosa¹, Dilza Balteiro Pereira de Campos¹, Isabela de Carvalho Leitão¹, R S Pinto¹, D Silva Dos Santos¹, Bruna Farjun¹, Dayana da Silva De Araújo¹, Fernanda Cristina Paccola Mesquita¹, G Monnerat-Cahli¹, E H Medei^{1

2

3}, Tais Hanae Kasai-Brunswick^{1

2

3}, A C Campos De Carvalho^{1

2

3

4}

Affiliations

¹ Carlos Chagas Filho Institute of Biophysics, Federal University of Rio de Janeiro, Rio de Janeiro, Brazil.
² National Center for Structural Biology and Bioimaging, Federal University of Rio de Janeiro, Rio de Janeiro, Brazil.
³ National Institute of Science and Technology in Regenerative Medicine, Rio de Janeiro, Brazil.
⁴ National Institute of Cardiology, Rio de Janeiro, Brazil.

Abstract

Human pluripotent stem cells (PSC) have been used for disease modelling, after differentiation into the desired cell type. Electrophysiologic properties of cardiomyocytes derived from pluripotent stem cells are extensively used to model cardiac arrhythmias, in cardiomyopathies and channelopathies. This requires strict control of the multiple variables that can influence the electrical properties of these cells. In this article, we report the action potential variability of 780 cardiomyocytes derived from pluripotent stem cells obtained from six healthy donors. We analyze the overall distribution of action potential (AP) data, the distribution of action potential data per cell line, per differentiation protocol and batch. This analysis indicates that even using the same cell line and differentiation protocol, the differentiation batch still affects the results. This variability has important implications in modeling arrhythmias and imputing pathogenicity to variants encountered in patients with arrhythmic diseases. We conclude that even when using isogenic cell lines to ascertain pathogenicity to variants associated to arrythmias one should use cardiomyocytes derived from pluripotent stem cells using the same differentiation protocol and batch and pace the cells or use only cells that have very similar spontaneous beat rates. Otherwise, one may find phenotypic variability that is not attributable to pathogenic variants.

Keywords: action potential (AP); cardiomyocytes; cell lines; differentiation batches; differentiation methods; healthy donors; iPSC (induced pluripotent stem cell); variability.