Tracheal tissue engineering in rats


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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Figure 1: Scaffolds for tissue-engineered rat trachea.
Figure 2: Biomechanical testing of rat tracheal scaffolds.
Figure 3: Two-photon imaging of live cells and LIVE/DEAD staining on the artificial graft 1 h after implantation.
Figure 4: Implantation of tissue-engineered tracheae in rats.


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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).

Author information

The concept of tracheal tissue engineering for both biological and artificial scaffolds was developed by P.M. P.J. and S.B. developed the decellularization concept for human trachea. J.C.H. and S.B. developed the decellularization concept for rodent trachea. The animal orthotopic transplantation model was designed by P.J. and conducted by P.J., J.C.H. and S.S. Cell seeding and scaffold evaluation was done by M.L.L., Y.G. and A.B.R. The mathematical model was developed by G.L. The mechanical evaluation setting was developed and conducted by C.D.G. and A.B. I.D. and P.U. were responsible for two-photon image acquisition and processing. The manuscript was approved by all coauthors before submission.

Correspondence to Paolo Macchiarini.

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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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