Table 2. Decellularization conditions.

	Treatment	Protocol with continuous agitation
1	Control	In PBS
2	Hyperosmotic	(i) 1.5 M NaCl in PBS (24 h) * # § ς
3	Ionic detergent	(i) 0.1% SDS in PBS (24 h) * # §
4	Non-ionic detergent	(i) 0.1% Tx in PBS (24 h) #
5	Ionic & non-ionic detergents	(i) 0.1% SDS(ii) 0.1% Tx in PBS (24 h) Φ
6	Enzymatic (2Nu)	(i) 1.5 M NaCl in PBS (24 h),(ii) 2 U/ml nuclease in NaCl/PBS (24 h, twice)
7	Enzymatic & detergent (2Nu + SDS)	(i) 1.5 M NaCl in PBS (24 h),(ii) 2 U/ml nuclease in NaCl/PBS (24 h, twice),(iii) 0.1% SDS in PBS (24 h)
8	Enzymatic (5Nu)	(i) 1.5 M NaCl in PBS (24 h),(ii) 5 U/ml nuclease in NaCl/PBS (24 h, twice) Φ ς
9	Enzymatic & detergent (5Nu + SDS)	(i) 1.5 M NaCl in PBS (24 h),(ii) 5 U/ml nuclease in NaCl/PBS (24 h, twice),(iii) 0.1% SDS in PBS (24 h)
10	Enzymatic (10Nu)	(i) 1.5 M NaCl in PBS (24 h),(ii) 10 U/ml nuclease in NaCl/PBS (24 h, twice),
11	Enzymatic & detergent (10Nu + SDS)	(i) 1.5 M NaCl in PBS (24 h),(ii) 10 U/ml nuclease in NaCl/PBS (24 h, twice),(iii) 0.1% SDS in PBS (24 h)

*Reference ²⁰; #Reference ³¹; §Reference ³⁴; ΦReference ²¹; ςReference ^22.

Note: NaCl, sodium chloride (Sigma-Aldrich); SDS, sodium dedecylsulfate (BioRad); Tx, Triton X-100 (Sigma); Nu, nuclease (DNase and RNase, Roche).