Collagenase & Dispase

is usually inefficient in dissociating tissues due to incomplete hydrolysis of all collagenous polypeptides and its limited activity against the high concentrations of non-collagen proteins and other macromolecules found in the extracellular matrix. For tissue dissociation, commonly used ‘Collagenases’ are collagenase preparations with additional enzymatic activities (side activity). They contain Collagenases, a Sulfhydryl Protease, Clostripain, a Trypsin-like Enzyme, and an Aminopeptidase. This combination of collagenolytic and proteolytic activities effectively breaks down intercellular matrices, particularly the peptide bonds present in native collagen.

Due to the varying compositions of extracellular matrices in different tissues and species, tailored collagenase preparations with optimized enzyme compositions have been developed. These are referred to as Type 1 to Type 7 for non-animal free products, and as Type A to Type D for the animal-free products. This classification helps determine the optimal enzyme mixture for the tissue type being used.

Explore the range of available Collagenase and Dispase® products,  and find a detailed table of their preparation speciality, and common application.

Dispase for Tissue Dissociation and Bioprocessing

Dispase®/Neutral Protease is an extremely stable non-specific metalloprotease which is produced by Paenibacillus polymyxa. Although related to trypsin, neutral protease is significantly less harmful to cells and can help prevent unwanted cell clumping without cell membrane damage. It cleaves fibronectin, collagen IV, and to a lesser extent collagen I, but it does not cleave collagen V or laminin. Thanks to its gentle proteolytic activity, this enzyme is particularly well-suited for preparing primary cells and for secondary cell culture (subcultivation), as it preserves cell membranes. Additionally, it serves as a secondary enzyme in cell isolation and tissue dissociation procedures, often in conjunction with Collagenases. It is commonly employed to separate the skin epidermis from the dermis while preserving epithelial sheets. Furthermore, it facilitates the isolation of stem cells, hepatocytes, and other cell types.

Animal Free (AF) Dispases® and Collagenases

AF Dispase and Collagenases, as well as optimized blends, are derived from cultures grown in medium completely devoid of animal-based components and designed for bioprocessing applications where introduction of potential animal derived pathogens must be prevented.

Find an overview of Preparations of Collagenase and Dispase®/Neutral Protease and their applications.

The choice and combination of enzymes depends on the tissue type, experimental requirements, and desired outcomes of the tissue dissociation procedure. The Worthingtons’ Tissue Dissociation Guide provides a collection of Tissue Dissociation Protocols for many different tissues from a broad spectrum of species.

DNase

DNase is also used in tissue dissociation and culture work to digest DNA from damaged cells thereby reducing viscosity, and removing membrane bound DNA fragments. Therefore, cell aggregation is prevented. Codes: DP and DCLS are suitable for these applications.