Human engineered skeletal muscle of hypaxial origin from pluripotent stem cells with advanced function and regenerative capacity

Human pluripotent stem cell derived muscle models show great potential for translational research. Here, we describe novel developmentally inspired methods for the derivation of skeletal muscle cells and their utility in three-dimensional skeletal muscle organoid formation as well as skeletal muscle tissue engineering. Three-dimensional organoid and tissue engineered models exhibit organotypic maturation and function and regenerative responses, recapitulating canonical properties of bona fide skeletal muscle in vivo. Key steps include the directed differentiation of human pluripotent stem cells to embryonic muscle progenitors of hypaxial origin followed by primary and secondary fetal myogenesis with development of a satellite cell pool and evidence for innervation in vitro. Regenerative competency was demonstrated in a cardiotoxin injury model with evidence for satellite cell activation as underlying mechanism. Overall design: Total RNA transcriptome of skeletal muscle differentiation of human pluripotent stem cells obtained at day 0, 1, 4, 8, 13, 22, 29, 60 of 2D differentiation and day 60 3D engineered skeletal muscle (ESM).