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Engineering Assembloids to Mimic Graft-Host Skeletal Muscle Interaction.

Lucia Rossi1,2, Beatrice Auletta1,2,3, Luigi Sartore1,2

  • 1Department of Molecular Medicine, University of Padova, Via G. Colombo 3, Padova, 35131, Italy.

Advanced Healthcare Materials
|May 5, 2025
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Summary
This summary is machine-generated.

This study developed a 3D human neuromuscular (NM) assembloid model using engineered skeletal muscle (SkM) and neuromuscular organoids (NMOs). The model successfully demonstrated functional neuromuscular junction (NMJ) formation and SkM regeneration after damage, aiding graft evaluation.

Keywords:
assembloidsmuscle regenerationmuscle stem cellneuromuscular organoidsorganoidsskeletal muscletissue engineering

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Area of Science:

  • Regenerative Medicine
  • Tissue Engineering
  • Neuroscience

Background:

  • Skeletal muscle (SkM) tissue engineering seeks to create implantable 3D constructs for muscle repair.
  • Evaluating the functional integration of engineered SkM with the host's innervated system is crucial for graft success.
  • A humanized in vitro model is needed to study neuromuscular interactions and graft potential.

Purpose of the Study:

  • To develop a 3D in vitro model for investigating the human neuromuscular (NM) system's response to engineered SkM.
  • To assess the function, stability, and adaptability of engineered SkM constructs within a human NM environment.
  • To create a tool for evaluating the graft potential of engineered SkM.

Main Methods:

  • Constructed 3D graft-host SkM assembloids using decellularized SkM (dSkM) as engineered tissue and human neuromuscular organoids (NMOs) as the recipient NM system.
  • Investigated cellular interactions, including myogenic cell migration and neural axon invasion from NMOs into dSkM constructs.
  • Assessed the formation of functional neuromuscular junctions (NMJs) and the regenerative capacity of the assembloids following induced damage.

Main Results:

  • Observed successful migration of myogenic cells and invasion of neural axons from NMOs into the engineered dSkM constructs.
  • Demonstrated the formation of functional neuromuscular junctions (NMJs) within the assembloids.
  • Showcased the ability of the assembloids to regenerate following acute damage, with SkM regeneration and functional recovery.

Conclusions:

  • The developed SkM assembloid model effectively mimics the human neuromuscular interaction between engineered SkM and an innervated system.
  • The model facilitates in vitro evaluation of neuromuscular junction formation, graft integration, and regenerative potential.
  • Despite limitations (lack of immune cells and vasculature), the assembloid is a valuable tool for assessing engineered SkM graft responses.