demonstrated that the single iPSC-CM micropatterning approach has advantages of single cell precision and control over cell shape, which was shown to exert a large influence on contractile function.( Ribeiro et al., 2015) Additionally, the single cell iPSC-CM method avoids interactions with non-myocytes, which are generally admixed into 3D iPSC-CM microtissues to maintain tissue integrity but may increase the potential for batch-to-batch heterogeneity.( Thavandiran et al., 2013) Multiple approaches have been reported to assess contractile function in iPSC-CMs, including motion assessment in 2D monolayers, 3D microtissue approaches, and the single cell micropatterning approach, as recently reviewed.( Blair and Pruitt, 2020) While 2D monolayer motion analysis is simple, disorganized myofibrillar organization limits accuracy and reproducibility. This analysis provides compelling evidence that myofibrillar structure should be assessed concurrently in studies investigating contractile function in iPSC-CMs.Ĭardiomyocytes derived from induced pluripotent stem cell (iPSC-CMs) have high potential as a model system for investigating causal mechanisms in cardiomyopathies and for pharmaceutical testing. Controlling for myofibrillar structure reduces false positive findings related to batch effect and improves sensitivity for pharmacologic testing and disease modeling. This variability is not reduced by subcloning from single iPSCs and is independent of iPSC-CM purification method. Automated analysis of micropatterned iPSC-CMs labeled with a cell permeant F-actin dye revealed that myofibrillar abundance is widely variable among iPSC-CMs and strongly correlates with contractile function. The objective of this study was to determine the impact of myofibrillar structure on contractile function in iPSC-CMs. ![]() ![]() Micropatterning iPSC-CMs on elastic substrates controls cell shape and alignment to enable contractile studies, but determinants of intrinsic variability in this system have been incompletely characterized. Disease modeling and pharmaceutical testing using cardiomyocytes derived from induced pluripotent stem cell (iPSC-CMs) requires accurate assessment of contractile function.
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