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Am J Transl Res 2010;2(1):43-55
Original Article
Engineering of aligned skeletal muscle by micropatterning
Ngan F. Huang, Randall J. Lee, Song Li,
Joint Graduate Program in Bioengineering, University of California Berkeley/University of California San
Francisco, Berkeley/San Francisco, CA, 94720, USA; Department of Medicine and the Cardiovascular Research
Institute, University of California San Francisco, San Francisco, CA; Department of Bioengineering, University of
California Berkeley, Berkeley, CA
Received August 7, 2009; accepted September 19, 2009; available online January 1, 2010
Abstract: Tissue engineered skeletal muscle has tremendous potential for the treatment of muscular injury or
muscular dysfunction. However, in vitro methods to generate skeletal muscle with physiologically aligned
myofiber structure remains limited. To develop a robust in vitro model that resembles the physiologically aligned
structure of muscle fibers, we fabricated micropatterned polymer membranes of poly(dimethylsiloxane) (PDMS)
with parallel microgrooves, and then examined the effect of micropatterning on myoblast cellular organization and
the cell fusion process. In comparison to the myoblasts on non-patterned PDMS films, myoblasts on
micropatterned PDMS films had well-organized F-actin assembly in close proximity to the direction of
microgrooves, along with enhanced levels of myotube formation at early time points. However, expression of
contractile proteins was not significantly affected by micropatterned substrates at the transcriptional and protein
levels. Similar results of cellular alignment was observed when myoblasts were cultured on microfluidically
patterned poly(2-hydroxyethyl methacrylate) (pHEMA) microgrooves. To apply this technology for generating
aligned tissue-like muscle constructs, we developed a methodology to transfer the aligned myotubes into
biodegradable collagen gels. Histological analysis revealed the persistence of aligned cellular organization in the
collagen gels. Together, these results demonstrate that topographically micropatterned PDMS or pHEMA can
promote cell alignment and fusion along the direction of the microgrooves, and this platform can be utilized to
transfer aligned myotubes on biodegradable hydrogels. This study highlights the importance of spatial cues in
creating aligned skeletal muscle for tissue engineering and muscular regeneration applications. (AJTR908001).
Key words: Micropatterning, myoblast, skeletal muscle, tissue engineering, fusion, alignment
Full Text PDF
Address all correspondence to:
Song Li, Ph.D.
University of California Berkeley
Department of Bioengineering
Berkeley, CA 94720-1762
Tel: (510) 666-2799, Fax: (510) 666-3381
Email: song_li@berkeley.edu
Original Article
Engineering of aligned skeletal muscle by micropatterning
Ngan F. Huang, Randall J. Lee, Song Li,
Joint Graduate Program in Bioengineering, University of California Berkeley/University of California San
Francisco, Berkeley/San Francisco, CA, 94720, USA; Department of Medicine and the Cardiovascular Research
Institute, University of California San Francisco, San Francisco, CA; Department of Bioengineering, University of
California Berkeley, Berkeley, CA
Received August 7, 2009; accepted September 19, 2009; available online January 1, 2010
Abstract: Tissue engineered skeletal muscle has tremendous potential for the treatment of muscular injury or
muscular dysfunction. However, in vitro methods to generate skeletal muscle with physiologically aligned
myofiber structure remains limited. To develop a robust in vitro model that resembles the physiologically aligned
structure of muscle fibers, we fabricated micropatterned polymer membranes of poly(dimethylsiloxane) (PDMS)
with parallel microgrooves, and then examined the effect of micropatterning on myoblast cellular organization and
the cell fusion process. In comparison to the myoblasts on non-patterned PDMS films, myoblasts on
micropatterned PDMS films had well-organized F-actin assembly in close proximity to the direction of
microgrooves, along with enhanced levels of myotube formation at early time points. However, expression of
contractile proteins was not significantly affected by micropatterned substrates at the transcriptional and protein
levels. Similar results of cellular alignment was observed when myoblasts were cultured on microfluidically
patterned poly(2-hydroxyethyl methacrylate) (pHEMA) microgrooves. To apply this technology for generating
aligned tissue-like muscle constructs, we developed a methodology to transfer the aligned myotubes into
biodegradable collagen gels. Histological analysis revealed the persistence of aligned cellular organization in the
collagen gels. Together, these results demonstrate that topographically micropatterned PDMS or pHEMA can
promote cell alignment and fusion along the direction of the microgrooves, and this platform can be utilized to
transfer aligned myotubes on biodegradable hydrogels. This study highlights the importance of spatial cues in
creating aligned skeletal muscle for tissue engineering and muscular regeneration applications. (AJTR908001).
Key words: Micropatterning, myoblast, skeletal muscle, tissue engineering, fusion, alignment
Full Text PDF
Address all correspondence to:
Song Li, Ph.D.
University of California Berkeley
Department of Bioengineering
Berkeley, CA 94720-1762
Tel: (510) 666-2799, Fax: (510) 666-3381
Email: song_li@berkeley.edu
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