Abstract
Directed differentiation methods allow acquisition of high-purity cardiomyocytes differentiated from human induced pluripotent stem cells (hiPSCs); however, their immaturity characteristic limits their application for drug screening and regenerative therapy. The rapid electrical pacing of cardiomyocytes has been used for efficiently promoting the maturation of cardiomyocytes, here we describe a simple device in modified culture plate on which hiPSC-derived cardiomyocytes can form three-dimensional self-organized tissue rings (SOTRs). Using calcium imaging, we show that within the ring, reentrant waves (ReWs) of action potential spontaneously originated and ran robustly at a frequency up to 4 Hz. After 2 weeks, SOTRs with ReWs show higher maturation including structural organization, increased cardiac-specific gene expression, enhanced Ca2+-handling properties, an increased oxygen-consumption rate, and enhanced contractile force. We subsequently use a mathematical model to interpret the origination, propagation, and long-term behavior of the ReWs within the SOTRs.
Highlights
Directed differentiation methods allow acquisition of high-purity cardiomyocytes differentiated from human induced pluripotent stem cells; their immaturity characteristic limits their application for drug screening and regenerative therapy
Fluorescence-activated cell sorting (FACS) (Supplementary Fig. 1b), voltage dye staining, and immunostaining data indicated that the self-organized tissue rings (SOTRs) were mostly ventricular cardiomyocytes and few fibroblast cells (Supplementary Figs. 3 and 4)
Previous studies promoted cardiomyocyte maturation by increasing their beating rates; the high beat rates of human induced pluripotent stem cells (hiPSCs)-derived cardiomyocytes could only be achieved by applying high-frequency electrical stimulation[3,14,16], making the sustainable application of such stimuli technically challenging
Summary
Directed differentiation methods allow acquisition of high-purity cardiomyocytes differentiated from human induced pluripotent stem cells (hiPSCs); their immaturity characteristic limits their application for drug screening and regenerative therapy. We subsequently use a mathematical model to interpret the origination, propagation, and long-term behavior of the ReWs within the SOTRs. human-induced pluripotent stem cell (hiPSC) derived cardiomyocytes have been proposed as an abundant resource for tissue engineering, drug screening, and regenerative-medicine applications[1], they exhibit characteristics different from adult human cardiomyocytes, including immature sarcomere structure and morphology, a fetus-like gene expression profile, and inadequate Ca2+-handling properties[2,3,4]. We create a platform capable of promoting rapid formation of hiPSC-derived cardiomyocytes into 3D selforganized tissue rings (SOTRs), where propagation of an action potential in the form of ReWs can spontaneously originate and travel around the closed-loop circuit, thereby making the cardiomyocytes beat at a high frequency (~2–4 Hz) continuously and robustly up to more than 89 days without any external stimulation. To our knowledge, a novel approach for spontaneous cardiomyocyte maturation and could serve as an economical and practical system for future production of mature hiPSC-derived cardiomyocytes
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