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. Author manuscript; available in PMC: 2012 Apr 1.
Published in final edited form as: Biomaterials. 2011 Feb 5;32(12):3233–3243. doi: 10.1016/j.biomaterials.2011.01.057

Table 2.

Selected agents and techniques for decellularizing tissue.

Agent/Technique Mode of action Effects on ECM References
Chemical Agents
Acids and bases Solubilizes cytoplasmic components of cells, disrupts nucleic acids, tend to denature proteins May damage collagen, GAG, and growth factors [18, 20–22, 35, 45, 46, 64, 65, 77, 97, 99, 103, 112]
Hypotonic and hypertonic solutions Cell lysis by osmotic shock, disrupt DNA-protein interactions Effectively lyses cells, but does not effectively remove cellular residues [20, 27, 29, 31, 41, 42, 45, 50, 51, 53, 54, 60, 67, 83, 97, 103, 108, 109]
Non-ionic detergents Disrupt DNA-protein interactions, disrupt lipid-lipid and lipid-protein interactions and to a lesser degree protein-protein interactions
 - Triton X-100 Mixed results with efficacy dependent on tissue, more effective cell removal from thin tissues, some disruption of ultrastructure and removal of GAG, less effective than SDS [8, 9, 11, 18, 20, 22, 25–29, 31–37, 41–43, 46, 48, 53, 54, 74, 77, 83, 99, 100, 109, 112]
Ionic detergents Solubilize cell and nucleic membranes, tend to denature proteins
 - Sodium dodecyl sulfate (SDS) Effectively removes nuclear remnants and cytoplasmic proteins from dense tissues, tends to disrupt ultrastructure, removes GAG and growth factors and damages collagen [8–11, 22–25, 28–36, 40, 41, 43, 45, 47, 53, 71, 73, 77, 100, 103, 107, 108]
 - Sodium deoxycholate Mixed results with efficacy dependent on tissue thickness, some disruption of ultrastructure and removal of GAG [18, 25, 26, 32–34, 40–42, 46, 48, 71, 74, 77, 83, 98, 99, 106, 109, 113]
 - Triton X-200 More effectively removes cells from thin tissues but with greater disruption of ultrastructure compared to other detergents [74]
Zwitterionic detergents Exhibit properties of non-ionic and ionic detergents
 - CHAPS Effectively removes cells with mild disruption of ultrastructure in thin tissues [10, 25, 73, 74]
 - Sulfobetaine-10 and -16 (SB-10, SB-16) Effectively removes cells with mild disruption of ultrastructure in thin tissues [74]
Solvents
 - Alcohols Cell lysis by dehydration, solubilize and remove lipids Effectively removes cells from dense dense tissues and inactivates pyrogens, but crosslinks and precipitates proteins, including collagen [21, 22, 28, 50, 51, 60, 65, 67, 75, 77, 80, 104]
 - Acetone Cell lysis by dehydration, solubilizes and removes lipids Effectively removes cells from dense dense tissues and inactivates pyrogens, but crosslinks and precipitates proteins, including collagen [28, 60, 65, 80]
 - Tributyl phosphate (TBP) Forms stable complexes with metals, disrupts protein-protein interactions Mixed results with efficacy dependent on tissue, dense tissues lost collagen but impact on mechanical properties was minimal [34–36]
Biologic Agents
Enzymes
 - Nucleases Catalyze the hydrolysis of ribonucleotide and deoxyribonucleotide chains Difficult to remove from the tissue, could invoke an immune response [10, 20, 23, 26, 29, 31, 33, 42, 44, 47, 50, 53, 67, 77, 83, 98, 99, 103, 106, 113]
 - Trypsin Cleaves peptide bonds on the C-side of Arg and Lys Prolonged exposure can disrupt ECM ultrastructure, removes ECM constituents such as collagen, laminin, fibronectin, elastin, and GAG, slower removal of GAG compared to detergents [18, 21, 22, 24, 26, 27, 29, 32, 33, 37, 38, 40, 41, 46, 48, 54, 77, 83]
 - Dispase Cleaves specific peptides, mainly fibronectin and collagen IV Prolonged exposure can disrupt ECM ultrastructure, removes ECM components such as fibronectin and collagen IV [21, 24, 39]
Chelating Agents (EDTA, EGTA) Chelating agents bind metallic ions, thereby disrupting cell adhesion to ECM Typically used with enzymatic methods (e.g. trypsin) but can be used with other agents, ineffective when used alone [18, 21–24, 26, 27, 29, 32, 33, 37–42, 45, 46, 48, 54, 73, 77, 83, 103]
Physical and Miscellaneous Agents
Temperature (freezing and thawing) Intracellular ice crystals disrupt cell membrane Ice crystal formation can disrupt or fracture ECM [18, 21, 23, 39, 46–48, 50, 89, 91, 92, 112, 117]
Direct application of force Removal of tissue eliminates cells and force can burst remaining cells Force can directly damage ECM [29, 39, 48, 51, 77, 97]
Pressure Pressure can burst cells and aid in removal of cellular material Pressure can disrupt ECM [33, 43, 44, 104]
Electroporation Pulsed electrical fields disrupt cell membranes Electrical field oscillation can disrupt ECM [95, 96]
Techniques to Apply Agents
Perfusion Facilitates chemical exposure and removal of cellular material Pressure associated with perfusion can disrupt ECM [8–10, 30, 46, 47, 53, 96, 99, 100]
Pressure gradient across tissue Facilitates chemical exposure and removal of cellular material Pressure gradient can disrupt ECM [21, 80, 103]
Supercritical fluid Pressure can burst cells, supercritical fluid facilitates chemical exposure and removal of cellular material Pressure necessary for supercritical phase can disrupt ECM [104]
Agitation Can lyse cells, but more commonly used to facilitate chemical exposure and removal of cellular material Aggressive agitation or sonication can disrupt ECM [11, 18, 22, 23, 25–29, 31, 32, 34–40, 42, 44, 48, 49, 52, 54, 58, 60, 71, 73, 74, 77, 80, 83, 89, 90, 97, 98, 103–113]