Table 3.

Adipose tissues. Decellularization techniques, biomechanical tests, recellularization methods, and in vivo implant of adipose extracellular matrix.

Tissues	Decellularization Methods	Biomechanical Tests	In Vitro Recellularization	In Vivo Implant	References
Subcutaneous adipose tissue from abdomen, breast, omentum, pericardial depot, thymic remnants	Freezing–thawing cycles + 0.25% trypsin and 0.1% EDTA + 99.9 2-propanol + 8 g/L NaCl, 200 mg/L KCl, 1 g/L Na2HPO4, 200 mg/L KH2PO4 + 55 mM Na2HPO4, 17 mM KH2PO4, 4.9 mM MgSO4, DNase, RNase, lipase	Indentation and compression tests	Static/dynamic seeding on ECM fragments/microcarriers/bead foams or encapsulation into hydrogel scaffolds of: -human adipose-derived stem cells -rat adipose-derived stem cells -human preadipocytes -chronic wound human fibroblasts	Rats and athymic mice: subcutaneous implantation	Flynn, 2010 [104] Turner and Flynn, 2012 [107] Turner et al., 2012 [108] Porzionato et al., 2013 [13] Yu et al., 2013 [112] Omidi et al., 2014 [102] Cheung et al., 2014 [113] Han et al., 2015 [114] Brown et al., 2015 [115] Baker et al., 2017 [103] Yu et al., 2017 [116] Shridhar et al., 2017 [117] Morissette Martin et al., 2018 [119]
	1% SDS + 2.5 mM sodium deoxycholate, lipase, and colipase	-	Seeding on hydrogel scaffolds of human adipose-derived stem cells	Athymic mice: injection of acellular adipose matrix in the subcutaneous dorsal region	Young et al., 2011 [106]
	2.5 mM sodium deoxycholate + lipase and colipase
	Mechanical agitation + 0.1, 1, 3, 5% peracetic acid + 1% Triton X-100 in 2 mM EDTA + 600 U/mL DNase	-	Seeding on hydrogel ECM of human adipose-derived stem cells	Rats: subcutaneous injection	Wu et al., 2012 [109]
Adipose tissue surgically sampled from abdomen, breast and forearm	Ultrasonic homogenization + 0.25% pancreatin	-	Culture of human adipose-derived stem cells with ECM microparticles	Nude mice: subcutaneous implantation of cell/scaffold complexes	Wang et al., 2013a [110]
	Freezing–thawing cycles + 0.5-1 M NaCl + 0.25% trypsin/EDTA + 1% Triton X-100	-	Seeding on adipose tissue microparticles of human adipose-derived stem cells	Nude rats: subcutaneous injection	Wang et al., 2013b [111]
	Decellularization according to Flynn, 2010 [104]	-	-	-	Sano et al., 2014 [126]
	Freezing–thawing cycles
	DNase + 4% SDS + 1% sodium azide
	1% Triton X-100 + DNase
	Freezing–thawing cycles + 0.25% trypsin/EDTA + 1% Triton X-100 + isopropanol	-	Seeding of breast cancer cells	-	Dunne et al., 2014 [127]
	1% SDS + 2.5 mM sodium deoxycholate + 100 µg/ml lipase and 50 ng/ml colipase	Compression tests	Encapsulation into the composite hydrogels of rat adipose-derived stem cells	Rats: subcutaneous tissue	Kayabolen et al., 2017 [118]
	Freezing–thawing cycles + 0.5% trypsin + 99.9% isopropanol + 500 U/mL Benzonase®	-	-	Mice: subcutaneous dorsal tissue	He et al., 2018 [120]
	Decellularization according to Flynn, 2010 [104]	-	-	GFP+ transgenic mice: subcutaneous tissue	Thomas-Porch et al., 2018 [121]
	1 M NaCl + 1 mM EDTA + Lysis buffer (1% tergitol type NP-40, 0.1% SDS, 5 mM EDTA, 0.4M NaCl, 50 mM Tris-HCl pH 8, 1 mM PMSF)
	Homogenization + 2 M urea buffer
Subcutaneous adipose tissue from cadavers	Organic solvent + surfactant/ethanol based solution	-	-	Immunocompetent and athimic mice: subcutaneous tissue	Giatsidis et al., 2018 [125]
Adipose tissue from cosmetic, abdominoplasty and liposuction surgery	Homogenization + freezing in 2 M urea buffer + 0.5% pepsin in 0.5 M acetic acid	Compression tests	Encapsulation into ECM-PEG hydrogels of human adipose-derived stem cells	Mice: subcutaneous implant of non-seeded ECM-PEG hydrogels	Li et al., 2018 [122]
	Freezing–thawing cycles + 99.9% isopropanol 0.05% trypsin/0.05% EDTA, 20 ng/mL DNase and 20 ng/ml RNase	Tensile tests	Seeding of human adipose-derived stem cells	-	Song et al., 2018 [123]
	Freezing–thawing cycles + homogenization + 1% Triton X-100 + 100 U/mL DNase and 100 µg/ml RNase	-	Encapsulation into ECM hydrogels of human adipose-derived stem cells	Rats: subcutaneous injection of unseeded hydrogel scaffolds	Zhao et al., 2018 [124]

SDS, sodium dodecyl sulfate; PEG, poly(ethylene glycol); DNase, deoxyribonuclease; RNase, ribonuclease; PMSF, phenylmethylsulfonyl fluoride; + means that separate cycles were performed; ‘and’ means that a mixture was performed between different substances.