Tissue-engineered tracheal transplants have been successfully performed clinically. However, before becoming a routine clinical procedure, further preclinical studies are necessary to determine the underlying mechanisms of in situ tissue regeneration. Here we describe a protocol using a tissue engineering strategy and orthotopic transplantation of either natural decellularized donor tracheae or artificial electrospun nanofiber scaffolds into a rat model. The protocol includes details regarding how to assess the scaffolds' biomechanical properties and cell viability before implantation. It is a reliable and reproducible model that can be used to investigate the crucial aspects and pathways of in situ tracheal tissue restoration and regeneration. The model can be established in <6 months, and it may also provide a means to investigate cell-surface interactions, cell differentiation and stem cell fate.
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This work was supported by European Project FP7-NMP-2011-SMALL-5: BIOtrachea, Biomaterials for Tracheal Replacement in Age-related Cancer via a Humanly Engineered Airway (no. 280584); by ALF medicine (Stockholm County Council): Transplantation of bio-engineered trachea in humans (no. LS1101–0042); by Vetenskapsrådet: Klinisk tillämpning av biokonstruerade organ med särskild betoning på trachea (No. K2013-99X-22252-01-5); by Dr. Dorka-Stiftung (Hannover, Germany): Bioengineering of tracheal tissue; and a Mega grant of the Russian Ministry of Education and Science (agreement no. 11.G34.31.0065).
The authors declare no competing financial interests.
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Jungebluth, P., Haag, J., Sjöqvist, S. et al. Tracheal tissue engineering in rats. Nat Protoc 9, 2164–2179 (2014) doi:10.1038/nprot.2014.149
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