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Published Online: 27 June 2014

Decellularization of Fibroblast Cell Sheets for Natural Extracellular Matrix Scaffold Preparation

Publication: Tissue Engineering Part C: Methods
Volume 21, Issue Number 1


The application of cell-derived extracellular matrix (ECM) in tissue engineering has gained increasing interest because it can provide a naturally occurring, complex set of physiologically functional signals for cell growth. The ECM scaffolds produced from decellularized fibroblast cell sheets contain high amounts of ECM substances, such as collagen, elastin, and glycosaminoglycans. They can serve as cell adhesion sites and mechanically strong supports for tissue-engineered constructs. An efficient method that can largely remove cellular materials while maintaining minimal disruption of ECM ultrastructure and content during the decellularization process is critical. In this study, three decellularization methods were investigated: high concentration (0.5 wt%) of sodium dodecyl sulfate (SDS), low concentration (0.05 wt%) of SDS, and freeze-thaw cycling method. They were compared by characterization of ECM preservation, mechanical properties, in vitro immune response, and cell repopulation ability of the resulted ECM scaffolds. The results demonstrated that the high SDS treatment could efficiently remove around 90% of DNA from the cell sheet, but significantly compromised their ECM content and mechanical strength. The elastic and viscous modulus of the ECM decreased around 80% and 62%, respectively, after the high SDS treatment. The freeze-thaw cycling method maintained the ECM structure as well as the mechanical strength, but also preserved a large amount of cellular components in the ECM scaffold. Around 88% of DNA was left in the ECM after the freeze-thaw treatment. In vitro inflammatory tests suggested that the amount of DNA fragments in ECM scaffolds does not cause a significantly different immune response. All three ECM scaffolds showed comparable ability to support in vitro cell repopulation. The ECM scaffolds possess great potential to be selectively used in different tissue engineering applications according to the practical requirement.

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Information & Authors


Published In

cover image Tissue Engineering Part C: Methods
Tissue Engineering Part C: Methods
Volume 21Issue Number 1January 2015
Pages: 77 - 87
PubMed: 24866751


Published in print: January 2015
Published ahead of print: 30 June 2014
Published online: 27 June 2014
Published ahead of production: 27 May 2014
Accepted: 7 May 2014
Received: 28 October 2013


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Qi Xing, PhD
Department of Biomedical Engineering, Michigan Technological University, Houghton, Michigan.
Keegan Yates
Department of Biomedical Engineering, Michigan Technological University, Houghton, Michigan.
Mitchell Tahtinen
Department of Biomedical Engineering, Michigan Technological University, Houghton, Michigan.
Emily Shearier, BSc
Department of Biomedical Engineering, Michigan Technological University, Houghton, Michigan.
Zichen Qian, BSc
Department of Biomedical Engineering, Michigan Technological University, Houghton, Michigan.
Feng Zhao, PhD
Department of Biomedical Engineering, Michigan Technological University, Houghton, Michigan.


Address correspondence to:Feng Zhao, PhDDepartment of Biomedical EngineeringMichigan Technological University1400 Townsend DriveHoughton, MI 49931E-mail: [email protected]

Disclosure Statement

The authors declare no competing financial interests.

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