Tissue Engineering of Skeletal Muscle

Loss of skeletal muscle profoundly affects the health and well-being of patients, and there currently is no way to replace lost muscle. We believe that a key step in the development of a prosthesis for reconstruction of dysfunctional muscular tissue is the ability to reconstitute the in vivo-like 3-dimensional (3D) organization of skeletal muscle in vitro with isolated satellite cells. In our present proof of principle studies, we have successfully constructed a multilayered culture of skeletal muscle cells, derived from neonatal satellite cells, that are distributed in a 3D pattern of organization that mimics many of the features of intact tissue. These multilayered cultures are composed of elongated multinucleated myotubes that are MyoD positive. Histological studies indicate that the multiple layers of myotubes can be distinguished. Expression of muscle-specific markers such as myosin heavy chain, dystrophin, integrin alpha-7, alpha-enolase, and beta-enolase was detected using real-time reverse transcriptase polymerase chain reaction at levels near adult values. Physiological measurements of the engineered skeletal muscle showed that they tetanize and display physiologic force length behavior, although developed force per cross-sectional area was below that of native rat skeletal muscle.

[1]  D J Mooney,et al.  Craniofacial tissue engineering. , 2001, Critical reviews in oral biology and medicine : an official publication of the American Association of Oral Biologists.

[2]  K. Davies,et al.  Distinct dystrophin mRNA species are expressed in embryonic and adult mouse skeletal muscle , 1988, FEBS letters.

[3]  M. Sillence,et al.  β2‐Agonist administration reverses muscle wasting and improves muscle function in aged rats , 2004, The Journal of physiology.

[4]  Cees W J Oomens,et al.  Monitoring local cell viability in engineered tissues: a fast, quantitative, and nondestructive approach. , 2003, Tissue engineering.

[5]  Caroline Joyce,et al.  Quantitative RT-PCR. A review of current methodologies. , 2002, Methods in molecular biology.

[6]  B. Nadal-Ginard,et al.  Characterization of a developmentally regulated perinatal myosin heavy-chain gene expressed in skeletal muscle. , 1984, The Journal of biological chemistry.

[7]  R. Bischoff,et al.  A satellite cell mitogen from crushed adult muscle. , 1986, Developmental biology.

[8]  R. Bischoff Enzymatic liberation of myogenic cells from adult rat muscle , 1974, The Anatomical record.

[9]  H. Vandenburgh,et al.  Tissue-engineered human bioartificial muscles expressing a foreign recombinant protein for gene therapy. , 1999, Human gene therapy.

[10]  M. Tanaka,et al.  Switching in levels of translatable mRNAs for enolase isozymes during development of chicken skeletal muscle. , 1985, Biochemical and biophysical research communications.

[11]  W. Song,et al.  H36-alpha 7 is a novel integrin alpha chain that is developmentally regulated during skeletal myogenesis [published erratum appears in J Cell Biol 1992 Jul;118(1):213] , 1992, The Journal of cell biology.

[12]  A. Minty,et al.  Sequential accumulation of mRNAs encoding different myosin heavy chain isoforms during skeletal muscle development in vivo detected with a recombinant plasmid identified as coding for an adult fast myosin heavy chain from mouse skeletal muscle. , 1983, Journal of Biological Chemistry.

[13]  T. Hawke,et al.  Myogenic satellite cells: physiology to molecular biology. , 2001, Journal of applied physiology.

[14]  Ellen M Arruda,et al.  Structure and functional evaluation of tendon-skeletal muscle constructs engineered in vitro. , 2006, Tissue engineering.

[15]  Y. Ide,et al.  Myosin Heavy Chain Isoforms of the Murine Masseter Muscle During Pre‐ and Post‐natal Development , 2003, Anatomia, histologia, embryologia.

[16]  H. Blau,et al.  Laminin-Induced Change in Conformation of Preexisting α7β1 Integrin Signals Secondary Myofiber Formation , 2001 .

[17]  K. Sakimura,et al.  cDNA cloning and nucleotide sequence of rat muscle‐specific enolase (ββ enolase) , 1989 .

[18]  A. Monaco,et al.  Conservation of the Duchenne muscular dystrophy gene in mice and humans. , 1987, Science.

[19]  T. Borg,et al.  Modulation of cardiac myocyte phenotype in vitro by the composition and orientation of the extracellular matrix , 1994, Journal of cellular physiology.

[20]  Robert G. Dennis,et al.  Excitability and isometric contractile properties of mammalian skeletal muscle constructs engineered in vitro , 2000, In Vitro Cellular & Developmental Biology - Animal.

[21]  M. Gilmore,et al.  Two-component regulator of Enterococcus faecalis cytolysin responds to quorum-sensing autoinduction , 2002, Nature.

[22]  A. Monaco,et al.  The complete sequence of dystrophin predicts a rod-shaped cytoskeletal protein , 1988, Cell.