3D skeletal muscle model

Scope of the method

The Method relates to
  • Animal health
  • Human health
The Method is situated in
  • Basic Research
  • Translational - Applied Research
Type of method
  • In vitro - Ex vivo
This method makes use of
  • Human derived cells / tissues / organs
Specify the type of cells/tissues/organs
myoblasts, fibroblasts, endothelial cells derived from human muscle tissue


Method keywords
  • Coculture
  • Myoblasts
  • endothelial cells
  • Bio-artificial muscle
  • Intramuscular injection
  • Compound testing
Scientific area keywords
  • tissue engineering
  • Muscle model
  • Skeletal muscle
  • Atrophy
  • Sarcopenia
  • Cachexia
  • Hypertrophy
  • Tissue development
  • mechanobiology
Method description

We tissue-engineer in vitro, skeletal muscle consisting of aligned myofibers. To create the so-called bio-artificial muscle (BAM), human muscle progenitor cells are expanded, and a 3D construct is created by mixing the cells with a hydrogel. The cell-gel mix is cast into custom-made silicone molds with end attachment sites and then differentiated for 1 week. The passive forces generated in the contracted hydrogel align the myogenic cells parallel to the long axis of the contracted gel such that they fuse into aligned multinucleated myofibers. This results in the formation of a 2 cm long and ~1.5 mm tick human BAM construct with endothelial networks. In addition, by co-culture with endothelial cells, interspersed endothelial networks can be created.

Lab equipment
  • - Incubator,
  • - Biology safety cabinet,
  • - Custom molds,
  • - Fluorescence microscope.
Method status
  • Published in peer reviewed journal

Pros, cons & Future potential


The model system allows for extensive biochemical, physical, cellular and electrical characterization of the effect of adding compounds, different extracellular matrix components or different cell types to investigate the effects on muscle development, morphology and function. It thus bridges the gap between 2D culture systems and in vivo experiments related to muscle tissue.

  • - Limited size of the constructs imposed by the limit of passive diffusion of nutrients and gases.
  • - Developmental stage of the muscle is comparable to foetal tissue, but can be stimulated to induce maturation (involves longer culture time).
  • - Integration with flow system, stimulation methods are under development.
  • - Cells derived from mouse, pig, rabbit, and cow muscles can also be used.
Future & Other applications

Besides further development toward regenerative medicine, such a muscle model can also be used to study mechanisms underlying myogenesis, vasculogenesis, and drug effects, administered either to the medium surrounding the muscle either by injection in the muscle.

References, associated documents and other information


Gholobova D., Decroix L., Van Mulyder V., Desender L., Gerard M., Carpentier G., Vandenburgh H. and Thorrez L. (2015) Endothelial network formation within human tissue-engineered skeletal muscle Tissue Eng. Part A 21 2548–58. doi.org/10.1089/ten.tea.2015.0093

Gholobova D., Gerard M., Terrie L., Desender L., Shansky J., Vandenburgh H., Thorrez L. Coculture Method to Obtain Endothelial Networks Within Human Tissue-Engineered Skeletal Muscle. Methods Mol Biol. 2019; 1889:169-183. doi: 10.1007/978-1-4939-8897-6_10

Gholobova, D. et al. Human tissue-engineered skeletal muscle: a novel 3D in vitro model for drug disposition and toxicity after intramuscular injection. Sci. Rep. 8, 12206 (2018)

Thorrez, L., DiSano, K., Shansky, J. & Vandenburgh, H. Engineering of Human Skeletal Muscle With an Autologous Deposited Extracellular Matrix. Front. Physiol. 9, 1–11 (2018)

Gholobova D., Terrie L., Mackova K., Desender L., Carpentier G., Gerard M., Hympanova L., Deprest J. and Thorrez L. Functional evaluation of prevascularization in one-stage versus two-stage tissue engineering approach of human bio-artificial muscle. Biofabrication 12 (2020) 035021. doi.org/10.1088/1758-5090/ab8f36

Research group muscles and movement

Contact person

Lieven Thorrez


Katholieke Universiteit Leuven (KUL)
Development and Regeneration
Research group muscles and movement
Flemish Region