dispase

Dispase is a neutral proteolytic enzyme used in cell biology to detach adherent cells and to separate epithelial tissues in laboratory workflows. It is produced by Bacillus species and is available in two main preparations, commonly referred to as Dispase I and Dispase II. The enzyme is favored for gentle tissue dissociation and the isolation of intact epithelial sheets.

Dispase acts by cleaving proteins in the basement membrane, notably collagen IV, laminin, and to a lesser extent fibronectin. This targeted activity allows the separation of epidermal or other epithelial layers from underlying connective tissue with relatively little damage to cell surface proteins and overall cell viability, in contrast to more aggressive proteases such as trypsin.

Common applications include the isolation of keratinocytes and other epithelial cells from skin and mucosal tissues, as well as the preparation of single-cell suspensions from cultured tissues for downstream analyses such as flow cytometry or culture. Dispase is typically used at neutral pH (around 7–8) and incubated at approximately 37°C for 20–60 minutes, after which proteolysis is halted by neutralization with serum-containing medium. Addition of DNase may be used to reduce cell clumping.

Limitations include batch-to-batch variability in activity, slower action compared with trypsin, and the potential for over-digestion if incubation is extended. Dispase can be more costly, and careful optimization of concentration and time is often required to preserve tissue integrity while achieving detachment.