Tissue Dissocation

Collagenases

Collagenase cleaves the peptide bonds in native, triple-helical collagen. Because of its unique ability to hydrolyze native collagen, it is widely used in isolation of cells from animal tissue. Collagenases occur in a variety of microorganisms and many different animal cells. The most potent collagenase is the “crude” collagenase secreted by the anaerobic bacterium Clostridium histolyticum. C. histolyticum collagenases have molecular weights from 68,000 to 125,000 Da and are metalloproteinases that require zinc and calcium. Our purified collagenase products have only trace amounts of caseinase (proteolytic) or clostripain activities. The purified Type VII Collagenase is also offered in cell culture tested and sterile-filtered versions. For additional information, please visit our collagenase, Dispase®, and Liberase® enzyme technical resource page.

Trypsin

Trypsin is a serine protease commonly used for detachment of adherent cell lines and dissociation of tissues. Crude trypsin preparations have typically been found to be more efficient for both applications. Cultured cells are most commonly removed from the culture substrate by treatment with trypsin, or trypsin-EDTA solutions. The concentration of trypsin can range from 0.025% to 0.5%. However, incubating cells with too high a trypsin concentration for too long a time period will damage cell membranes and kill the cells. For the dissociation of tissues, trypsin has been used alone or as a supplement to other enzymes. For additional information, please visit our Trypsin technical resources page.

Additional Tissue Dissociation and Cell Detachment Enzymes

Our comprehensive portfolio contains a large variety of tissue dissociation enzymes for your specific cell culture or tissue dissociation needs. For example, hyaluronidase is typically used as a supplement to proteases for tissue dissociation. It catalyzes the random hydrolysis of 1,4-b-D-glycosidic linkages between N-acetylgalactosamine or N-acetylgalactosamine sulfate and glucuronic acid in hyaluronic acid, chondroitin, chondroitin 4- and 6-sulfates, and dermatan. Various nucleases, such as DNase and RNase help to reduce viscosity resulting from nucleic release from damaged cells during harvesting. Papain is a relatively nonspecific sulfhydryl protease derived from papaya latex. Papain is used alone or in addition to other proteases such as collagenase. For additional information, explore our enzymes and technical resources for your tissue dissociation and cell detachment workflow needs.