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Table 1 Experimental details for the five different decellularization protocols

From: Preparation and characterization of small-diameter decellularized scaffolds for vascular tissue engineering in an animal model

Protocol step Group
I II III IV V
Soaking in ddw for 24 h
Frozen at −80 °C and thawed twice
Soaking: 75% ethanol
Solvent:tissue ratio: 20:1 (v/w)
Total duration: 72 h
Solvent replacement at 1, 3, 6, 12, 24, 72 h
Soaking in ddw for 24 h
Digestion on an orbital shaker using a tissue:enzyme ratio (g/ml) of 1:15 0.125% pepsin 0.125% pepsin 0.125% pepsin 0.125% pepsin 0.125% pepsin
Solvent 1× EDTA 2× EDTA 1× EDTA 1× EDTA 2× EDTA
Duration (h) 1.5 1 2 2 1.5
PBS washing for 30 min; shaking
DNase and RNase for 6 h; shaking
Rinsing solution ddw ddw Triton ddw Triton
Duration (h) 1 1 2 1 2
Ultraviolet irradiation for 30 min    
  1. DNase and RNase concentration was 70 U/ml, respectively
  2. Each step was performed at 4 °C
  3. n = 5 for each group, repeated six times in all
  4. ddw double-distilled water, Triton 1% Triton X100