Decellularized adipose matrix provides an inductive microenvironment for stem cells in tissue regeneration

Table 1 Different studies of decellularization and sterilization methods for preparation of DAM

Decellularization methods			Sterilization methods	Refs.
Physical treatments	Chemical treatments	Biological treatments
Freeze–thaw, 3 cycles (-80 °C to 37 °C)	99.9% isopropanol	0.25% trypsin/0.1% EDTA 15000 U DNase, 12.5 mg RNase, 2000 U lipase	70% ethanol/1% penicillin and streptomycin/UV light/1% antibiotic/antimycotic	[26,30,74,75,83,84,96,101,102]
		0.05% trypsin-EDTA 100 U/mL benzonase	70% ethanol/1% penicillin and streptomycin	[63,66]
		0.05% trypsin 500 U/mL benzonase	0.1% peracetic acid in 4% ethanol	[61]
	99.9% isopropanol 1 mol/L NaCl	1 mmol/L EDTA + Lysis buffer (1% tergitol type NP-40, 0.1% SDS, 5 mmol/L EDTA, 0.4 mol/L NaCl, 50 mmol/L Tris-HCl pH 8, 1 mmol/L PMSF)	70% ethanol/1% penicillin and streptomycin	[65]
	100% isopropanol	0.25% trypsin-EDTA; 1 mL DNase + 1 mL RNase + 2 mL lipase	70% ethanol/1% penicillin and streptomycin	[135]
	0.5 mol/L NaCl/1 mol/L NaCl/isopropanol/Triton X-100	0.25% trypsinEDTA	1% penicillin and streptomycin	[69,71,93]
Freeze-thaw, 3 cycles (-80 °C to 37 °C) + ultrasonic	0.5% SDS + 100% ethanol	—	100% ethanol	[25]
Freeze-thaw, 35 cycles (-80 °C to 37 °C) + homogenization, 5 min (12000 r/min)	1% Triton X100 + 100% isopropanol + 1 mol/L NaCl	100 U/mL DNase 100 μg/mL RNase	—	[68]
Freezethaw, 4 cycles (-80 °C to 37 °C) + ultrasonic	96% ethanol 0.5% SDS	0.05% trypsin/0.05 mmol/L EDTA + DNase	—	[87]
Freezethaw, 5 cycles (-80 °C to 37 °C)	Isopropanol	0.25% trypsin/0.1% EDTA DNase I + RNase A	Ethylene oxide	[136]
Freeze-thaw, 5 cycles (liquid nitrogen to 37 °C)	99.9% isopropanol	0.05% trypsin-EDTA 20 ng/mL DNase I + 20 ng/mL RNase	1% penicillin and streptomycin	[67]
Freezethaw, 4 or 5 cycles (liquid nitrogen to Room temperature)	0.5 mol/L acetic acid	—	—	[88]
Freezethaw, 35 cycles (liquid nitrogen to Room temperature)	0.1% SDS	0.05% trypsin + 0.05% EDTA + 20 ng/mL DNase I + 20 ng/mL RNase	1% penicillin and streptomycin	[89]
Freezethaw, 618 cycles (liquid nitrogen to room temperature)	0.1% sodium azide + 1 mol/L NaCl + 4% sodium deoxycholate	2000 K units DNase	—	[78]
	1% Triton X-100	2000 K units DNase
Homogenization, 5 min (12000 rpm)	1 mol/L NaCl/0.5% SDS	0.2% DNase + 200 μg/mL RNase	—	[27,28,81]
	—	—	—	[104,137]
	0.5% SDS + 100% isopropanol	—	—	[138]
Homogenization, 5 min + ultrasonic	—	0.25% Pancreatin	Ethylene oxide	[79]
Homogenization, 3 min (12000 r/min)	SDS	—	—	[90]
	4 mol/L urea	4 mol/L Gu	Ethylene oxide	[100,139]
Homogenization (twice)	2 mol/L urea+70% ethanol	2 U/mL dispase II + 4 mol/L GuHCl	Dialysis against chloroform	[23,24]
Homogenization	2 mol/L urea buffer	—	70% ethanol/1% antibiotic/antimycotic solution	[65]
Constant stirring	1% SDS or 2.5 mmol/L sodium deoxycholate	2.5 mmol/L sodium deoxycholate + 500 U porcine lipase + 500 U porcine colipase	365 nm UV light	[62,85]
Constant stirring	1% SDS	2.5 mmol/L sodium deoxycholate + 100 μg/mL lipase + 50 ng/mL colipase; 50 μg/mL DNase + 50 μg/mL RNase	Ethylene oxide	[86]
Mechanical processing	0.1%, 1%, 3%, or 5% Peracetic acid + 1% Triton X-100	600 U DNase	—	[31,94]
	3% Triton X100 + 4% sodium deoxycholate + 4% ethanol/0.1% peracetic acid + 100% n-propanol	0.02% trypsin + 0.05% EDTA	4% ethanol + 0.1% peracetic acid	[22]
SCCO2 (180 bar)	Ethanol	—	SC-CO2	[91]
—	1% sodium dodecylsulfate + 100% isopropanol	2.5 mmol/L sodium deoxycholate + 500 U lipase + 500 U colipase	5000 IU penicillin and 5 mg/mL streptomycin	[32]
	0.5% SDS + isopropanol + 0.1% peracetic acid + 4% ethanol	—	0.1% peracetic acid+4% ethanol	[92,103]
	1% Triton X-100	10, 20 and 100 IU/mL DNase I	—	[82]
	1-propanol	Sodium deoxycholate	Peracetic acid	[33]
	Organic solvent + surfactant/ethanol-based solution	—	Peracetic acid	[34,64]

SDS: Sodium dodecyl sulfate; EDTA: Ethylenediaminetetraacetic acid; SC-CO2: Supercritical carbon dioxide; Gu: guanidine.