Table 1.

Decellularization protocol.

1. Aortic allograft harvest

2. Wash with Hank’s (pH7.4) at 4°C

3. Rinse in hypotonic Tris-NaCl (0.05 mol/L NaCl, 0.02% EDTA, 0.05 mol/L Tris-Cl, 100kIU/ml aprotinin, pH8.0) at 4°C for 24 hours

4. Rinse in hypertonic Tris-NaCl (1.5 mol/L NaCl, 0.02% EDTA, 0.05 mol/L Tris-Cl, 100kIU/ml aprotinin, pH8.0) at 4°C for 24 hours

5. Wash with Hank’s (pH7.4) at 4°C, 15 minutes for 4 times

6. Rinse in Tris- Triton X − 100 (1%Triton X-100, 0.02% EDTA, 0.05 mol/L Tris-Cl, 100kIU/ml aprotinin, pH7.4) at 4°C for 24 hours

7. Wash with Hank’s (pH7.4 without Mg2+, Ca2+) at 4°C

8. Enzyme extraction: Rinse in Tris- Enzyme (10 mg/L DNAase I, 1 mg/L RNAaseA, 3 mmol/L Mg2+, 1 mmol/L Ca2+, 10 mmol/L Tris-Cl) at 37°C for 1–2 hours

9. Repeat step 6

10. Rinse in Hank’s (pH7.4) at 4°C for 48 hours