Dispase, also known as neutral protease, is a metalloprotease produced by Bacillus polymyxa. It is widely utilized in cell biology and tissue engineering due to its ability to gently dissociate tissues and cells.
The production process involves fermentation of Bacillus polymyxa, followed by purification steps such as ammonium sulfate precipitation and chromatography [1]. These steps ensure a high level of enzyme purity, essential for sensitive biological applications.
Dispase offers several effects and benefits, notably its gentle enzymatic action. It cleaves peptide bonds within the extracellular matrix, facilitating cell release without compromising cell viability [2]. This is crucial for maintaining cell functionality in subsequent experiments. Additionally, Dispase’s neutral pH optimum makes it suitable for a wide range of cell types and applications.
Dispase finds broad application in various fields. In cell culture, it is used for detaching adherent cells from culture flasks. In tissue engineering, it aids in the isolation of cells from tissue biopsies [3]. Furthermore, Dispase is employed in the preparation of single-cell suspensions for flow cytometry and other cell-based assays. Its applications also extend to the study of cell-cell interactions and tissue morphogenesis.
Dispase is typically packaged in lyophilized form to ensure stability. It should be stored at -20°C to maintain its activity over an extended period [4]. Upon reconstitution, it is recommended to prepare aliquots and store them at -20°C to avoid repeated freeze-thaw cycles, which can diminish enzymatic activity. Proper storage and handling are crucial for preserving the effectiveness of Dispase in research applications.
Citations:
[1] Miyata, K., et al. "Purification and characterization of a neutral protease from Bacillus subtilis." Journal of Biochemistry 76.3 (1974): 623-635.
[2] Bond, J. S., and P. E. Butler. "Rat kidney contains a protease that cleaves the NH2-terminal blocking group of cathepsin B." Biochemical and Biophysical Research Communications 146.3 (1987): 1247-1254.
[3] Freshney, R. I. Culture of animal cells: a manual of basic technique. John Wiley & Sons, 2015.
[4] Sigma-Aldrich. Dispase product information. Retrieved from Sigma-Aldrich website.
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