APPENDIX
Table 1
Commonly used decellularization methods and chaotropic agents
Method
Mode of action
Effects on ECM
Physical
Snap Freezing
Mechanical force
Mechanical agitation
Intracellular ice crystals disrupt
ECM can be disrupted or fractured
cell membrane
during rapid freezing
Pressure can burst cells and tissue
Mechanical force can cause damage to
removal eliminates cells
ECM
Can cause cell lysis, but more
Aggressive agitation or sonication can
commonly used to facilitate
disrupt ECM as the cellular material is
chemical exposure and
removed
cellular material removal
Chemical
Alkaline; acid
Solubilizes cytoplasmic
Removes GAGs (glycosaminoglycans)
components of cells; disrupts
nucleic acids
Non-ionic detergents
Disrupts lipid–lipid and lipid–
Mixed results; efficiency dependent on
Triton X-100
protein interactions, while leaving
tissue, removes GAGs
protein–protein interactions intact
Ionic detergents
Solubilize cytoplasmic and
Removes nuclear remnants and
Sodium dodecyl
nuclear cellular membranes; tend
cytoplasmic proteins; tends to disrupt
sulfate (SDS)
to denature
native tissue structure, remove GAGs
proteins
and damage collagen.
Sodium deoxycholate
More disruptive to tissue structure than
SDS
Triton X-200
Yielded efficient cell removal when
used with zwitterionic detergents
Zwitterionic detergents
CHAPS
Exhibit properties of non-ionic
Efficient cell removal with ECM
and ionic
disruption similar to that of Triton X-
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