Biofabrication of 3D Human Muscle Model with Vascularization and Endomysium

Methods Mol Biol. 2022;2373:213-230. doi: 10.1007/978-1-0716-1693-2_13.

Abstract

This protocol describes the biofabrication of 3D millimeter-scale human muscle units, embedding non-planar muscle fibers wrapped by fibroblasts-rich endomysium and intertwined with microvascular networks. Suspended muscle fibers are formed through the self-assembly of human myoblasts within cylindrical cavities generated in a sacrificial gelatin template cast in a 3D printed frame. Following myotube differentiation, muscle fibers are embedded in a 3D matrix containing endothelial cells and muscle-derived fibroblasts. The cellular complexity of the environment is instrumental to drive fibroblast migration towards muscle fibers and to induce the organ-specific differentiation of endothelial cells. This advanced 3D muscle model can be applied to analyze the biological mechanisms underlying specific muscle diseases which involve a complex remodeling of the muscle environment (e.g., muscular dystrophies and fibrosis) whereby the pathological interplay among different cell populations drives the onset and progression of the disease.

Keywords: 3D human muscle model; 3D printing; Duchenne muscular dystrophy; Endomysium; Endothelial cell; Fibrosis; Mesoscale; Organ specificity; Vasculature.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Endothelial Cells*
  • Humans
  • Muscle Fibers, Skeletal
  • Muscular Dystrophy, Duchenne
  • Myoblasts
  • Myocardium*