Table 1. Step-wise representation of the established protocol for the decellularization of porcine urethras.

Step	Time (h)	Conditions	Aim
Urethra dissection	0.5	N/A	Remove adipose tissue
Urethra cannulation	0.2	14 G cannula and surgical suture	Fix the cannula to one extremity of the urethra
Mount decellularization apparatus	0.2	Supplementary Figure S2	Attach the cannula to the perfusion tubing system
Wash urethra	0.5	Distilled water: 40 ml/min + 60 rpm	Wash out blood and debris
Add detergent solution	0.1	Detergent solution (details in Methods)	Replace distilled water by detergent solution
Start decellularization process	48	40 ml/min + 60 rpm	Remove cellular components of the tissue and preserve the ECM
Change perfusion direction	0.2	Change the cannula from one extremity of the urethra to the other	Allow both extremities to be equally exposed to the perfusion forces
Continue decellularization process	72	40 ml/min + 60 rpm	Remove cellular components of the tissue and preserve the ECM
Wash bioscaffold	≥ 12	Distilled water: 40 ml/min + 60 rpm	Remove detergent remnants
Total	≥ 133.7

N/A: not applicable.