Xing Q, Yates K, Tahtinen M, Shearier E, Qian Z, Zhao F, et al.
Tissue engineering. Part C, Methods. Date of publication 2015 Jan 1;volume 21(1):77-87.
1. Tissue Eng Part C Methods. 2015 Jan;21(1):77-87. doi: 10.1089/ten.TEC.2013.0666.
Decellularization of fibroblast cell sheets for natural extracellular matrix
scaffold preparation.
Xing Q(1), Yates K, Tahtinen M, Shearier E, Qian Z, Zhao F.
Author information:
(1)Department of Biomedical Engineering, Michigan Technological University ,
Houghton, Michigan.
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.
DOI: 10.1089/ten.tec.2013.0666
PMCID: PMC4291209
PMID: 24866751 [Indexed for MEDLINE]
