Protocol to develop force-generating human skeletal muscle organoids

STAR Protoc. 2024 Mar 15;5(1):102794. doi: 10.1016/j.xpro.2023.102794. Epub 2023 Dec 20.


Force generation is an essential property of skeletal muscle models in vitro. We describe a versatile 1-step procedure to direct undifferentiated human pluripotent stem cells (PSCs) into contractile skeletal muscle organoids (SMOs). Our protocol provides detailed steps for 3D casting of PSCs using either collagen-I/Matrigel- or fibrin/Geltrex-based hydrogels, SMO differentiation, and application of different culture platforms for mechanical loading and contractility analysis. The SMO model may be particularly useful to study human muscle development and developmental skeletal muscle disorders in vitro. For complete details on the use and execution of this protocol, please refer to Shahriyari et al.1.

Keywords: Cell Differentiation; Cell culture; Cell-based Assays; Organoids; Tissue Engineering.

MeSH terms

  • Cell Differentiation
  • Humans
  • Muscle, Skeletal
  • Organoids*
  • Pluripotent Stem Cells*