Annals of Biomedical Engineering

, Volume 44, Issue 6, pp 2076–2089 | Cite as

Understanding the Role of ECM Protein Composition and Geometric Micropatterning for Engineering Human Skeletal Muscle

  • Rebecca M. Duffy
  • Yan Sun
  • Adam W. Feinberg
Emerging Trends in Biomaterials Research


Skeletal muscle lost through trauma or disease has proven difficult to regenerate due to the challenge of differentiating human myoblasts into aligned, contractile tissue. To address this, we investigated microenvironmental cues that drive myoblast differentiation into aligned myotubes for potential applications in skeletal muscle repair, organ-on-chip disease models and actuators for soft robotics. We used a 2D in vitro system to systematically evaluate the role of extracellular matrix (ECM) protein composition and geometric patterning for controlling the formation of highly aligned myotubes. Specifically, we analyzed myotubes differentiated from murine C2C12 cells and human skeletal muscle derived cells (SkMDCs) on micropatterned lines of laminin compared to fibronectin, collagen type I, and collagen type IV. Results showed that laminin supported significantly greater myotube formation from both cells types, resulting in greater than twofold increase in myotube area on these surfaces compared to the other ECM proteins. Species specific differences revealed that human SkMDCs uniaxially aligned over a wide range of micropatterned line dimensions, while C2C12s required specific line widths and spacings to do the same. Future work will incorporate these results to engineer aligned human skeletal muscle tissue in 2D for in vitro applications in disease modeling, drug discovery and toxicity screening.


Skeletal muscle Tissue engineering Extracellular matrix Microcontact printing 



Extracellular matrix





Col I

Collagen I

Col IV

Collagen IV


Skeletal muscle derived cells


Microcontact printed


Myotube fusion index (nuclei/myotube)





Financial support from the National Institutes of Health Director’s New Innovator Award (DP2HL117750) to Adam Feinberg and the John and Claire Bertucci Fellowship from the Carnegie Institute of Technology to Rebecca Duffy. The lab of Johnny Huard at the University of Pittsburgh graciously provided initial aliquots of Cook Myosite human SkMDC and Cook MyoSite Inc. provided assistance with human SkMDC lot selection. We thank Lucas Friedman for providing assistance with image analysis.

Supplementary material

10439_2016_1592_MOESM1_ESM.pdf (2.6 mb)
Supplementary material 1 (PDF 2626 kb)


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Copyright information

© Biomedical Engineering Society 2016

Authors and Affiliations

  • Rebecca M. Duffy
    • 1
  • Yan Sun
    • 1
    • 2
  • Adam W. Feinberg
    • 1
    • 3
  1. 1.Regenerative Biomaterials and Therapeutics Group, Department of Biomedical EngineeringCarnegie Mellon UniversityPittsburghUSA
  2. 2.Key Laboratory for Biomechanics and Mechanobiology of Ministry of Education, School of Biological Science and Medical EngineeringBeihang UniversityBeijingChina
  3. 3.Department of Materials Science and EngineeringCarnegie Mellon UniversityPittsburghUSA

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