Skip to main content
SpringerLink
Log in

Mouse Skeletal Muscle Decellularization

  • Protocol
  • First Online:
Decellularized Scaffolds and Organogenesis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1577))

Abstract

Natural acellular matrices obtained from decellularization procedures are biocompatible and non-immunogenic materials considered promising tools for regenerative medicine purposes. Before in vivo implantation, these matrices must be efficiently decellularized, removing all the cellular components to avoid any immunogenic reaction. At the same time, it is important to maintain the original three-dimensional structure of the specific tissue. Here we describe a method: (1) to decellularize mouse quadriceps using a detergent-enzymatic treatment (DET) and (2) to assess decellularization efficiency and scaffold properties.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution

Institutional subscriptions

References

  1. Badylak SF (2004) Xenogeneic extracellular matrix as a scaffold for tissue reconstruction. Transpl Immunol 12:367–377. doi:10.1016/j.trim.2003.12.016

    Article  CAS  PubMed  Google Scholar 

  2. Daley WP, Peters SB, Larsen M (2008) Extracellular matrix dynamics in development and regenerative medicine. J Cell Sci 121:255–264. doi:10.1242/jcs.006064

    Article  CAS  PubMed  Google Scholar 

  3. Badylak SF, Taylor D, Uygun K (2011) Whole-organ tissue engineering: decellularization and recellularization of three-dimensional matrix scaffolds. Annu Rev Biomed Eng 13:27–53. doi:10.1146/annurev-bioeng-071910-124743

    Article  CAS  PubMed  Google Scholar 

  4. Gilbert TW, Sellaro TL, Badylak SF (2006) Decellularization of tissues and organs. Biomaterials 27:3675–3683. doi:10.1016/j.biomaterials.2006.02.014

    Article  CAS  PubMed  Google Scholar 

  5. Piccoli M, Urbani L, Alvarez-Fallas ME, Franzin C, Dedja A, Bertin E, Zuccolotto G, Rosato A, Pavan P, Elvassore N, De Coppi P, Pozzobon M (2016) Improvement of diaphragmatic performance through orthotopic application of decellularized extracellular matrix patch. Biomaterials 74:245–255. doi:10.1016/j.biomaterials.2015.10.005

    Article  CAS  PubMed  Google Scholar 

  6. Gillies AR, Smith LR, Lieber RL, Varghese S (2011) Method for decellularizing skeletal muscle without detergents or proteolytic enzymes. Tissue Eng Part C Methods 17:383–389. doi:10.1089/ten.TEC.2010.0438

    Article  CAS  PubMed  Google Scholar 

  7. Kasukonis BM, Kim JT, Washington TA, Wolchok JC (2016) Development of an infusion bioreactor for the accelerated preparation of decellularized skeletal muscle scaffolds. Biotechnol Prog 32:745–755. doi:10.1002/btpr.2257

    Article  CAS  PubMed  Google Scholar 

  8. Fishman JM, Lowdell MW, Urbani L, Ansari T, Burns AJ, Turmaine M, North J, Sibbons P, Seifalian AM, Wood KJ, Birchall MA, De Coppi P (2013) Immunomodulatory effect of a decellularized skeletal muscle scaffold in a discordant xenotransplantation model. Proc Natl Acad Sci U S A 110:14360–14365. doi:10.1073/pnas.1213228110

    Article  PubMed  PubMed Central  Google Scholar 

  9. Elliott MJ, De Coppi P, Speggiorin S, Roebuck D, Butler CR, Samuel E, Crowley C, McLaren C, Fierens A, Vondrys D, Cochrane L, Jephson C, Janes S, Beaumont NJ, Cogan T, Bader A, Seifalian AM, Hsuan JJ, Lowdell MW, Birchall MA (2012) Stem-cell-based, tissue engineered tracheal replacement in a child: a 2-year follow-up study. Lancet 380:994–1000. doi:10.1016/S0140-6736(12)60737-5

    Article  PubMed  PubMed Central  Google Scholar 

  10. Macchiarini P, Jungebluth P, Go T, Asnaghi MA, Rees LE, Cogan TA, Dodson A, Martorell J, Bellini S, Parnigotto PP, Dickinson SC, Hollander AP, Mantero S, Conconi MT, Birchall MA (2008) Clinical transplantation of a tissue-engineered airway. Lancet 372:2023–2030. doi:10.1016/S0140-6736(08)61598-6

    Article  PubMed  Google Scholar 

  11. Wainwright DJ (1995) Use of an acellular allograft dermal matrix (AlloDerm) in the management of full-thickness burns. Burns 21:243–248

    Article  CAS  Google Scholar 

  12. Sicari BM, Agrawal V, Siu BF, Medberry CJ, Dearth CL, Turner NJ, Badylak SF (2012) A murine model of volumetric muscle loss and a regenerative medicine approach for tissue replacement. Tissue Eng Part A 18:1941–1948. doi:10.1089/ten.TEA.2012.0475

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Sicari BM, Rubin JP, Dearth CL, Wolf MT, Ambrosio F, Boninger M, Turner NJ, Weber DJ, Simpson TW, Wyse A, Brown EH, Dziki JL, Fisher LE, Brown S, Badylak SF (2014) An acellular biologic scaffold promotes skeletal muscle formation in mice and humans with volumetric muscle loss. Sci Transl Med 6:234ra258. doi:10.1126/scitranslmed.3008085

    Article  CAS  Google Scholar 

  14. Crapo PM, Gilbert TW, Badylak SF (2011) An overview of tissue and whole organ decellularization processes. Biomaterials 32:3233–3243. doi:10.1016/j.biomaterials.2011.01.057

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgments

Piccoli M and Franzin C are supported by Istituto di Ricerca Pediatrica Città della Speranza, Grant Number 16/02; Trevisan C and Maghin E are supported by Istituto di Ricerca Pediatrica Città della Speranza. Pozzobon M is supported by University of Padova, Grant number GRIC15AIPF, Assegno di ricerca Senior. Franzin C, Piccoli M, and Pozzobon M are coinventors of Italian Patent N. 0001422436 entitled “Matrice acellulare per ricostruzione in vivo di muscolo scheletrico.”

Author information

Authors and Affiliations

  1. Fondazione Istituto di Ricerca Pediatrica Città della Speranza, Padova, Italy

    Martina Piccoli, Caterina Trevisan, Edoardo Maghin, Chiara Franzin & Michela Pozzobon

  2. Department of Women and Children Health, University of Padova, Padova, Italy

    Caterina Trevisan & Michela Pozzobon

  3. Stem Cells and Regenerative Medicine Lab, Istituto di Ricerca Pediatrica Città della Speranza, Corso Stati Uniti 4, 35127, Padova, Italy

    Michela Pozzobon

Authors
  1. Martina Piccoli
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Caterina Trevisan
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Edoardo Maghin
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Chiara Franzin
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Michela Pozzobon
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Michela Pozzobon .

Editor information

Editors and Affiliations

  1. Ottawa Hospital Research Institute, Ottawa, ON, Canada

    Kursad Turksen

Rights and permissions

Reprints and permissions

Copyright information

© 2017 Springer Science+Business Media New York

About this protocol

Cite this protocol

Piccoli, M., Trevisan, C., Maghin, E., Franzin, C., Pozzobon, M. (2017). Mouse Skeletal Muscle Decellularization. In: Turksen, K. (eds) Decellularized Scaffolds and Organogenesis. Methods in Molecular Biology, vol 1577. Humana Press, New York, NY. https://doi.org/10.1007/7651_2017_28

Download citation

  • DOI: https://doi.org/10.1007/7651_2017_28

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7655-3

  • Online ISBN: 978-1-4939-7656-0

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation