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

Abstract

SummaryHuman pluripotent stem cell derived muscle models show great potential for translational research. Here, we describe developmentally inspired methods for derivation of skeletal muscle cells and their utility in three-dimensional skeletal muscle organoid formation as well as skeletal muscle tissue engineering. 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 into hypaxial muscle with development of a satellite cell pool and evidence for innervation in vitro. Skeletal muscle organoids faithfully recapitulate all steps of embryonic myogenesis in 3D Tissue engineered muscle exhibits organotypic maturation and function, advanced by thyroid hormone. Regenerative competence was demonstrated in a cardiotoxin injury model with evidence of satellite cell activation as underlying mechanism. Collectively, we introduce a hypaxial muscle model with canonical properties of bona fide skeletal muscle in vivo to study muscle development, maturation, disease, and repair.

The Evolution of Complex Muscle Cell In Vitro Models to Study Pathomechanisms and Drug Development of Neuromuscular Disease

“…Regeneration: Several rodents and human models have demonstrated that functional regeneration of skeletal muscle by satellite cells is recapitulated in vitro [ 86 , 134 , 158 , 229 ]. It would be important to extend these findings to neuromuscular disease entities to dissect if particular mutations or disease states affect muscle stem cell function in an experimental setting.…”

“…Human induced pluripotent stem cells (iPSC) are another valuable source of skeletal myocytes. A number of efficient directed differentiation protocols have been introduced to derive skeletal myocyte from iPSC [ 82 , 83 , 84 , 85 , 86 , 87 ]. All of them aim to recapitulate the embryonic development of skeletal muscle in cell culture by temporal modification and simulation of developmental cues.…”

“…Tissue engineering of 3D muscle has been introduced more than 30 years ago by Vandenburgh et al [ 146 ]. This approach was recognized early on as a method for durable culturing of muscle cells [ 146 ] with reported culture durations of up to 3 months [ 86 ]. In addition, 3D muscle engineering may not only improve the cellular morphology of muscle cells but rather allows force measurements similar to primary muscle strips from patients [ 147 ].…”

Multiscale X-ray Structural Analysis of Cardiac Cells and Tissues

“…However, due to the immature state of the cells and the small X-ray spot size, the diffraction signal of the actomyosin lattice was quite low [226]. Nonetheless, progress in development of EHM patches of more mature cells gives hope for a detailed structural analysis [337,338]. Furthermore, the structural changes of the sarcomeric structure caused by genetic diseases such as Duchenne Muscle dystrophy could be investigated in a follow up experiment.…”