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Comparison of decellularization techniques for preparation of extracellular matrix scaffolds derived from three‐dimensional cell culture
Author(s) -
Lu Hongxu,
Hoshiba Takashi,
Kawazoe Naoki,
Chen Guoping
Publication year - 2012
Publication title -
journal of biomedical materials research part a
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.849
H-Index - 150
eISSN - 1552-4965
pISSN - 1549-3296
DOI - 10.1002/jbm.a.34150
Subject(s) - decellularization , extracellular matrix , tissue engineering , scaffold , materials science , biomedical engineering , plga , self healing hydrogels , matrix (chemical analysis) , biophysics , chemistry , nanotechnology , biochemistry , polymer chemistry , biology , composite material , medicine , nanoparticle
Extracellular matrix (ECM) scaffolds derived from cultured cells have drawn increasing attention for use in tissue engineering. We have developed a method to prepare cultured cell‐derived ECM scaffolds by combining three‐dimensional cell culture, decellularization, and selective template removal. Cell‐ECM‐template complexes were first formed by culture of cells in a poly(lactic‐ co ‐glycolic acid) (PLGA) mesh template to deposit their own ECM. The complexes were subsequently decellularized to remove cellular components. Finally, the PLGA template was selectively removed to obtain the ECM scaffolds. Seven decellularization methods were compared for their decellularization effects during scaffold preparation. They were: freeze‐thaw cycling (–80°C, six times) with ammonia water (25 m M ); 0.1% Triton™ X‐100 (TX100) with 1.5 M KCl aqueous solution; freeze‐thaw cycling alone; ammonia water alone; TX100 extraction; osmotic shock with 1.5 M KCl; and freeze‐thaw cycling with 3 M NaCl. Among these methods, the methods of freeze‐thaw cycling with NH 4 OH and TX100 with 1.5 M KCl showed the best effect on the removal of cellular components from the complexes, while the other five methods could only partially remove cellular components. The ECM scaffolds prepared by these two methods had similar gross appearances and microstructures. In vivo implantation of the ECM scaffolds prepared by these two methods induced mild host responses. The two decellularization methods were demonstrated to be effective for preparation of cultured cell‐derived ECM scaffolds. © 2012 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 100A: 2507–2516, 2012.