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

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.

Hydrogel‐Sheathed hiPSC‐Derived Heart Microtissue Enables Anchor‐Free Contractile Force Measurement

“…In drug screening using tissues derived from iPSCs of patients, even the slightest differences between tissues are important because of the large individual differences in cells. [ 19 ] Therefore, fiber‐shaped HMT has the advantage of high sensitivity and early testing because the number of cells used is small due to microscale tissue. Furthermore, considering the diffusion coefficient, small EHT is preferable for drug administration because the diffusion velocity of soluble drug is proportional to the square of the distance in the tissue.…”

Optical redox imaging to screen synthetic hydrogels for stem cell-derived cardiomyocyte differentiation and maturation

“…Efficiency of Batches in the Cardiac Progenitor Stage Confirming batch purity of stem cell-derived CMs is often labor-intensive, requiring techniques such as flow cytometry or qPCR that are typically performed two weeks after initializing differentiation. 3,8,9,63 Label-free FLIM of metabolic coenzymes can separate stem cell-derived CMs by differentiation efficiency within 24 hours of initializing differentiation. 46 We investigated if widefield ORI provides sufficient sensitivity to separate cell batches by differentiation efficiency in the CPC stage.…”

“…5,6 Stem cell-derived CMs also exhibit batch-to-batch and line-to-line variability in function and structure. [7][8][9][10][11] Furthermore, methods to assess differentiation and maturation efficiency [e.g., ribonucleic acid (RNA) sequencing, immunostaining] are generally destructive and low-throughput. 12,13 The efficient production and widespread use of iPSC-CMs will require consistent manufacturing of functionally mature cell batches and touchfree, higher throughput, and nondestructive monitoring.…”

Revolutionizing Disease Modeling: The Emergence of Organoids in Cellular Systems

“…These variations can be influenced by the methods of reprogramming, the quality of the starting cells, and the culture conditions used to maintain the iPSCs. This variability can make it difficult to compare results between studies and to predict the behavior of iPSC-derived cells in a clinical setting [ 63 , 64 , 65 , 66 ].…”