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Method for decellularizing skeletal muscle without detergents or proteolytic enzymes.

Publication ,  Journal Article
Gillies, AR; Smith, LR; Lieber, RL; Varghese, S
Published in: Tissue Eng Part C Methods
April 2011

Decellularized skeletal muscle is a promising model that can be used to study cell-matrix interactions and changes that occur in muscle extracellular matrix (ECM) in myopathies and muscle wasting diseases. The goal of this study is to develop a novel method to decellularize skeletal muscle that maintains the native biochemical composition and structure of the ECM. This method consists of sequential incubation of mouse tibialis anterior muscles in latrunculin B, high ionic strength salt solution, and DNase I and avoids use of proteases or detergents that degrade the ECM. Characterization of the decellularized muscles using hematoxylin and eosin staining along with DNA quantification suggested complete removal of DNA, whereas biochemical analyses indicated no loss of collagens and only a slight reduction in glycosaminoglycans. Western blot analysis of decellularized tissues showed removal of the vast majority of the contractile proteins actin and myosin, and morphological analysis using scanning electron microscopy suggested removal of myofibers from decellularized muscle tissues. Passive mechanical testing of decellularized muscle bundles revealed the typical nonlinear behavior, similar to that of intact muscle. Together, these results suggest that the protocol developed successfully decellularizes skeletal muscle without altering its composition and mechanical function.

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Published In

Tissue Eng Part C Methods

DOI

EISSN

1937-3392

Publication Date

April 2011

Volume

17

Issue

4

Start / End Page

383 / 389

Location

United States

Related Subject Headings

  • Tissue Engineering
  • Thiazolidines
  • Staining and Labeling
  • Peptide Hydrolases
  • Osmolar Concentration
  • Muscle, Skeletal
  • Mice, Inbred C57BL
  • Mice
  • Extracellular Matrix
  • Detergents
 

Citation

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Gillies, A. R., Smith, L. R., Lieber, R. L., & Varghese, S. (2011). Method for decellularizing skeletal muscle without detergents or proteolytic enzymes. Tissue Eng Part C Methods, 17(4), 383–389. https://doi.org/10.1089/ten.TEC.2010.0438
Gillies, Allison R., Lucas R. Smith, Richard L. Lieber, and Shyni Varghese. “Method for decellularizing skeletal muscle without detergents or proteolytic enzymes.Tissue Eng Part C Methods 17, no. 4 (April 2011): 383–89. https://doi.org/10.1089/ten.TEC.2010.0438.
Gillies AR, Smith LR, Lieber RL, Varghese S. Method for decellularizing skeletal muscle without detergents or proteolytic enzymes. Tissue Eng Part C Methods. 2011 Apr;17(4):383–9.
Gillies, Allison R., et al. “Method for decellularizing skeletal muscle without detergents or proteolytic enzymes.Tissue Eng Part C Methods, vol. 17, no. 4, Apr. 2011, pp. 383–89. Pubmed, doi:10.1089/ten.TEC.2010.0438.
Gillies AR, Smith LR, Lieber RL, Varghese S. Method for decellularizing skeletal muscle without detergents or proteolytic enzymes. Tissue Eng Part C Methods. 2011 Apr;17(4):383–389.

Published In

Tissue Eng Part C Methods

DOI

EISSN

1937-3392

Publication Date

April 2011

Volume

17

Issue

4

Start / End Page

383 / 389

Location

United States

Related Subject Headings

  • Tissue Engineering
  • Thiazolidines
  • Staining and Labeling
  • Peptide Hydrolases
  • Osmolar Concentration
  • Muscle, Skeletal
  • Mice, Inbred C57BL
  • Mice
  • Extracellular Matrix
  • Detergents