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. 2023 Oct 4;32:98–123. doi: 10.1016/j.bioactmat.2023.09.017

Table 1.

Physical, Enzymatic, and Chemical Methods commonly applied to decellularize tissues and organs.

Category
 Treatment/Technique
 Effect on Tissue
 Ref.
Decellularization Method
Chemical
Acids & Bases

  • -Causes/catalyzes hydrolytic degradation of biomolecules and solubilizing cytoplasmic components

 [252]
Hypotonic/Hypertonic Solutions

  • -Causes cell lysis due to induced osmotic effects

  • -Most often combined with other decellularized agents

 [253,254]
Detergents

  • -Destroys lipid-lipid & lipid-protein interactions, solubilizing plasma membranes of cells and dissociating genetic material from proteins
Non-ionic Detergents (e.g. Triton X-100)

  • -Non-denaturing detergent that causes cell lysis and removal of cellular residues

  • -Limited in ability to break protein-protein bonds

 [254,255]
Ionic Detergents (e.g. SDS)

  • -Modifies protein structures due to charge difference

  • -Disrupts cell membranes

 [256,257]
Zwitterionic Detergents (e.g. CHAPS)

  • -Non-denaturing detergent with properties of both ionic and non-ionic detergents

  • -Milder agents with greater ECM structural maintenance

  • -Incomplete cytoplasmic removal

 [118]
Enzymatic
Nucleases (DNase, RNase, etc.)

  • -Catalyzes the cleavage of phosphodiester bonds of nucleotides, fragmenting DNA and RNA for inactivation, prevention of replication and elimination

 [258]
Trypsin

  • -Cleaves peptide bonds of arginine and lysine

  • -Commonly used in combination with chelating agents such as ethylenediaminetetraacetic acid (EDTA) used to bind to divalent cations at cell-adhesion sites to cause dissociation

 [259,260]
Lipase

  • -Catalyzes the cleavage of ester bonds in lipids

 [261,262]
Dispase

  • -Protease that mainly cleaves fibronectin and collagen IV proteins

 [263,264]
Collagenase

  • -Cleaves peptide bonds in collagen

 [265,266]
Physical
Freeze-Thaw Cycling

  • -Can consist of one or more freeze-thaw cycles

  • -Lyses cell membrane via ice crystal formation

  • -Range of temperature −80 °C – 37 °C

 [[267], [268], [269], [270], [271]]
High Hydrostatic Pressure

  • -Relatively quick decellularization process

  • -High pressure disrupts cell membrane inside tissue

 [[272], [273], [274], [275], [276]]