Method for decellularizing skeletal muscle without detergents or proteolytic enzymes

Allison R. Gillies, Lucas R. Smith, Richard L. Lieber, Shyni Varghese

Research output: Contribution to journalArticle

70 Citations (Scopus)

Abstract

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.

Original languageEnglish (US)
Pages (from-to)383-389
Number of pages7
JournalTissue Engineering - Part C: Methods
Volume17
Issue number4
DOIs
StatePublished - Apr 1 2011

Fingerprint

Detergents
Muscle
Skeletal Muscle
Peptide Hydrolases
Muscles
Extracellular Matrix
Wasting Syndrome
Contractile Proteins
Deoxyribonuclease I
DNA
Muscular Diseases
Hematoxylin
Myosins
Eosine Yellowish-(YS)
Glycosaminoglycans
Tissue
Cell Communication
Electron Scanning Microscopy
Osmolar Concentration
Actins

ASJC Scopus subject areas

  • Bioengineering
  • Medicine (miscellaneous)
  • Biomedical Engineering

Cite this

Method for decellularizing skeletal muscle without detergents or proteolytic enzymes. / Gillies, Allison R.; Smith, Lucas R.; Lieber, Richard L.; Varghese, Shyni.

In: Tissue Engineering - Part C: Methods, Vol. 17, No. 4, 01.04.2011, p. 383-389.

Research output: Contribution to journalArticle

Gillies, Allison R. ; Smith, Lucas R. ; Lieber, Richard L. ; Varghese, Shyni. / Method for decellularizing skeletal muscle without detergents or proteolytic enzymes. In: Tissue Engineering - Part C: Methods. 2011 ; Vol. 17, No. 4. pp. 383-389.
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