Engineered skeletal muscle recapitulates human muscle development, regeneration and dystrophy

Mina Shahriyari, Md Rezaul Islam, Sadman M. Sakib, Malte Rinn, Anastasia Rika, Dennis Krüger, Lalit Kaurani, Verena Gisa, Mandy Winterhoff, Harithaa Anandakumar, Orr Shomroni, Matthias Schmidt, Gabriela Salinas, Andreas Unger, Wolfgang A. Linke, Jana Zschüntzsch, Jens Schmidt, Rhonda Bassel-Duby, Eric N. Olson, André FischerWolfram Hubertus Zimmermann, Malte Tiburcy

Research output: Contribution to journalArticlepeer-review

12 Scopus citations


Background: Human pluripotent stem cell-derived muscle models show great potential for translational research. Here, we describe developmentally inspired methods for the derivation of skeletal muscle cells and their utility in skeletal muscle tissue engineering with the aim to model skeletal muscle regeneration and dystrophy in vitro. Methods: Key steps include the directed differentiation of human pluripotent stem cells to embryonic muscle progenitors followed by primary and secondary foetal myogenesis into three-dimensional muscle. To simulate Duchenne muscular dystrophy (DMD), a patient-specific induced pluripotent stem cell line was compared to a CRISPR/Cas9-edited isogenic control line. Results: The established skeletal muscle differentiation protocol robustly and faithfully recapitulates critical steps of embryonic myogenesis in two-dimensional and three-dimensional cultures, resulting in functional human skeletal muscle organoids (SMOs) and engineered skeletal muscles (ESMs) with a regeneration-competent satellite-like cell pool. Tissue-engineered muscle exhibits organotypic maturation and function (up to 5.7 ± 0.5 mN tetanic twitch tension at 100 Hz in ESM). Contractile performance could be further enhanced by timed thyroid hormone treatment, increasing the speed of contraction (time to peak contraction) as well as relaxation (time to 50% relaxation) of single twitches from 107 ± 2 to 75 ± 4 ms (P < 0.05) and from 146 ± 6 to 100 ± 6 ms (P < 0.05), respectively. Satellite-like cells could be documented as largely quiescent PAX7+ cells (75 ± 6% Ki67) located adjacent to muscle fibres confined under a laminin-containing basal membrane. Activation of the engineered satellite-like cell niche was documented in a cardiotoxin injury model with marked recovery of contractility to 57 ± 8% of the pre-injury force 21 days post-injury (P < 0.05 compared to Day 2 post-injury), which was completely blocked by preceding irradiation. Absence of dystrophin in DMD ESM caused a marked reduction of contractile force (−35 ± 7%, P < 0.05) and impaired expression of fast myosin isoforms resulting in prolonged contraction (175 ± 14 ms, P < 0.05 vs. gene-edited control) and relaxation (238 ± 22 ms, P < 0.05 vs. gene-edited control) times. Restoration of dystrophin levels by gene editing rescued the DMD phenotype in ESM. Conclusions: We introduce human muscle models with canonical properties of bona fide skeletal muscle in vivo to study muscle development, maturation, disease and repair.

Original languageEnglish (US)
Pages (from-to)3106-3121
Number of pages16
JournalJournal of Cachexia, Sarcopenia and Muscle
Issue number6
StatePublished - Dec 2022


  • Duchenne muscular dystrophy
  • hypaxial dermomyotome
  • limb muscle
  • satellite cells
  • skeletal muscle organoid
  • somite
  • tissue engineering

ASJC Scopus subject areas

  • Orthopedics and Sports Medicine
  • Physiology (medical)


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