Dispase

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Introductions

Dispase is a protease that cleaves fibronectin, collagen IV and, to a lesser extent, collagen I. It is present in some bacteria and can be isolated from culture filtrates of polymyxa. It can be extracted, purified, and used for research. It is particularly useful for isolating embryonic epithelial cells and mesenchyme. Dispase II is specifically used to cleave leucine-phenylalanine bonds.

Properites

Dispase is a neutral protease, which is a non-specific metalloprotease used for digestion of extracellular matrix from different tissues or organs. Dispase can also be used for cell harvesting or inoculation transfer in cell culture. It is also used to prevent accidental clumping of cells in suspension cell culture. The commonly used trypsin, collagenase and streptase proteases are all protein hydrolases and are used to disperse tissues and separate cells, but they are highly injurious to cells, destabilize cells in culture, and even introduce mycoplasma contamination. Dispase does not harm cell membranes and does not introduce mycoplasma or any animal viruses. Differences in temperature, pH, serum composition, etc. have little effect on dispase, whose activity is greatly reduced by dilution, which also facilitates subsequent culture.

Enzyme Activity Definition

1U is the amount of enzyme that releases Folin-positive amino acids or peptides equivalent to 1 μmol of tyrosine for 1 min under assay conditions. Dispase: ≥ 0.8 U/mg (37 degrees, using casein as substrate, pH 7.5).

Inhibitors: EDTA, EGTA, Hg²⁺. Serum does not inhibit enzyme activity.

Advantages

• Rapid and effective yet gentle action of the enzyme.
• The cells can remain intact without damage to the cell membrane.
• The enzyme is of bacterial origin and contains no mycoplasma or animal virus contamination.
• Stable to changes in temperature, pH, and serum composition.
• Simple inactivation with chelating agents or dilution.

Applications

• Dispase is often used to digest adherent primary cells in culture, as this treatment turns out to be gentler than trypsin digestion.
• Dispase can digest serine-phenylalanine.
• Dispase intravitreal injection can be used to model proliferative vitreoretinal lesions in different animals.
• Dispase is a class of neutral metalloprotein hydrolases that can recycle cells grown on matrigel substrates. In addition, dispase can also be used for tissue separation.