Circulating re-entrant waves promote maturation of hiPSC-derived cardiomyocytes in self-organized tissue ring

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 hiPSCderived 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 Ca 2+-handling properties, an increased oxygenconsumption 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.

Phenotypic recapitulation and correction of desmoglein-2-deficient cardiomyopathy using human-induced pluripotent stem cell-derived cardiomyocytes

“…1B ). We recently developed a simple device in modified culture plate on which iPSC-CMs form three-dimensional (3D) SOTRs that show higher maturation, including structural organization ( 33 ). To investigate the structural strength and force generation of the organized tissue lacking desmoglein-2 expression, we generated SOTRs from the isogenic iPSC-CMs and evaluated their contractile force using a MicroTester G2.…”

Precision 3D‐Printed Cell Scaffolds Mimicking Native Tissue Composition and Mechanics

“…The immortalized mouse myoblast cell line (C2C12) is a widely used model to study the biomechanics of single cells [70,71] and differentiating tissues [72] on substrates that mimic the ECM in natural conditions of myocytes (YM ≈12 kPa). [73] Regulation and maintenance of muscle mass is facilitated by the differentiation of skeletal muscles to myotubes and an optimal muscle function is accompanied by the alignment of actin filaments. C2C12 can proliferate, form confluent tissues and differentiate into myotubes under the right cell culture conditions in vitro.…”

hiPSC-Derived Cardiac Tissue for Disease Modeling and Drug Discovery

“…The 3D structure facilitates CMs self-organizing into an advanced and complex structure. Li et al developed a device based on a low-attachment substrate where hiPSC-CMs can self-organize into a 3D tissue ring [ 138 , 139 ]. Without any external stimulation, the tissue ring cultured in the 3D system could spontaneously generate re-entrant waves, accompanied by rapid pacing, thus leading to enhanced maturation of CMs in an autonomous manner.…”