l

Trachea positioning: 5–8 min. This time may be vari-

able, the trachea is submerged in medium, therefore it

could take as long as needed.

(d)

Motor and pump installation: 10 min.

(e)

Tracheal graft de-epithelialization: 5 h.

(f)

Graft decontamination: 36–48 h.

Acknowledgments

This work was supported in part of the University of Toronto’s

Medicine by Design initiative, which receives funding from the

Canada First Research Excellence Fund (CFREF; C1TPA-2016-

18).

References

1. Gilbert TW, Sellaro TL, Badylak SF (2006)

Decellularization of tissues and organs. Bioma-

terials 27:3675–3683

2. Scarrit ME, Pachos NC, Bunnell BA (2015) A

review of cellularization strategies for tissue

engineering of whole organs. Front Bioeng

Biotechnol 3:1–17

3. Remlinger NT, Czajka CA, Juhas ME et al

(2010) Hydrated xenogeneic decellularized

tracheal matrix as a scaffold for tracheal recon-

struction. Biomaterials 31:3520–3526

4. Batioglu-Karaaltin A, Karaaltin MV, Ovali E

et al (2015) In vivo tissue-engineered allogenic

trachea transplantation in rabbits: a preliminary

report. Stem Cell Rev Rep 11:347–356

5. Delaere P, Van Raemdonck D (2016) Tracheal

replacement. J Thorac Dis 8:S186–S196

6. Delaere P, Vranckx J, Verleden G et al (2010)

Tracheal allotransplantation after withdrawal of

immunosuppressive therapy. N Engl J Med

362:138–145

7. Delaere PR (2012) Tracheal transplantation.

Curr Opin Pulm Med 18:313–320

8. Aoki FG, Varma R, Marin-Araujo AE et al

(2019) De-epithelialization of porcine tracheal

allografts in a bioreactor system: implications in

tissue engineering approaches for

tracheal

replacement. Sci Rep 9:12034

9. Lee H, Marin-Araujo AE, Aoki FG et al (2021)

Computational fluid dynamics for enhanced

tracheal bioreactor design and long-segment

graft recellularization. Sci Rep 11:1187

10. Haykal S, Salna M, Zhou Y et al (2014)

Double-chamber

rotating

bioreactor

for

dynamic

perfusion

cell

seeding

of

large-

segment tracheal allografts: comparison to con-

ventional static methods. Tissue Eng Part C

Methods 20:8

11. Łopianiak I, Butruk-Raszeja BA (2020) Evalu-

ation of sterilization/disinfection methods of

fibrous polyurethane scaffolds designed for tis-

sue engineering applications. Int J Mol Sci

21:8092

12. JoVE Science Education Database (2021) Lab

animal research, sterile tissue harvest protocol.

JoVE,

Cambridge,

MA.

https://www.jove.

com/v/10298/sterile-tissue-harvest.

Accessed 12 Jun 2021

182

Alba E. Marin-Araujo et al.