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Published 13 September 2004. doi:10.1083/jcb.200405004
The Rockefeller University Press, 0021-9525 $8.00
JCB, Volume 166, Number 6, 877-887
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Article

 Myotubes differentiate optimally on substrates with tissue-like stiffness : pathological implications for soft or stiff microenvironments



Adam J. Engler1, Maureen A. Griffin1, Shamik Sen1, Carsten G. Bönnemann2, H. Lee Sweeney2,3, and Dennis E. Discher1,2,3

1 School of Engineering and Applied Science, University of Pennsylvania, Philadelphia, PA 19104
2 Pennsylvania Muscle Institute, University of Pennsylvania, Philadelphia, PA 19104
3 Cell and Molecular Biology Graduate Group, University of Pennsylvania, Philadelphia, PA 19104

Address correspondence to Dennis E. Discher, Cell and Molecular Biology Graduate Group, University of Pennsylvania, 112 Towne Building, 220 S. 33rd St., Philadelphia, PA 19104. Tel.: (215) 898-4809. Fax: (215) 573-6334. email: discher{at}seas.upenn.edu

Contractile myocytes provide a test of the hypothesis that cells sense their mechanical as well as molecular microenvironment, altering expression, organization, and/or morphology accordingly. Here, myoblasts were cultured on collagen strips attached to glass or polymer gels of varied elasticity. Subsequent fusion into myotubes occurs independent of substrate flexibility. However, myosin/actin striations emerge later only on gels with stiffness typical of normal muscle (passive Young's modulus, E ~12 kPa). On glass and much softer or stiffer gels, including gels emulating stiff dystrophic muscle, cells do not striate. In addition, myotubes grown on top of a compliant bottom layer of glass-attached myotubes (but not softer fibroblasts) will striate, whereas the bottom cells will only assemble stress fibers and vinculin-rich adhesions. Unlike sarcomere formation, adhesion strength increases monotonically versus substrate stiffness with strongest adhesion on glass. These findings have major implications for in vivo introduction of stem cells into diseased or damaged striated muscle of altered mechanical composition.

Key Words: myofibrillogenesis; patterning; adhesion; differentiation; muscular dystrophy

Abbreviations used in this paper: AFM, atomic force microscopy; EDL, extensor digitorum longus; IPN, interpenetrating polymer network; PA, polyacrylamide; SMC, smooth muscle cell.


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