What is Dispase?
Dispase is a neutral, proteolytic enzyme derived from the bacterium Bacillus polymyxa. It is commonly used in histology and cell culture to gently dissociate tissue into single cells without damaging cell membranes or extracellular matrix components.
How Does Dispase Work?
Dispase functions by cleaving fibronectin, collagen IV, and to a lesser extent, collagen I. This selective cleavage allows for the detachment of cells from the extracellular matrix while preserving surface proteins and cell viability.
Applications in Histology
Dispase is widely used in various histological applications: Tissue dissociation: Dispase is used to separate epithelial layers from underlying stroma, making it easier to study specific cell types.
Cell isolation: It is employed to isolate primary cells from tissues, such as keratinocytes from skin or endothelial cells from blood vessels.
Organoid culture: Dispase helps in generating organoids by dissociating cells from tissues, which can then self-organize into 3D structures.
Immunohistochemistry: It aids in the preparation of samples by dissociating tissues into single cells, which can then be used for antibody staining and analysis.
Advantages of Using Dispase
Dispase offers several advantages for histological applications: Gentle dissociation: It minimizes cell damage and maintains cell surface markers, making it ideal for downstream applications like flow cytometry or cell sorting.
High viability: Cells isolated using Dispase generally exhibit high viability and functionality, which is crucial for accurate experimental results.
Versatility: It is suitable for a wide range of tissues and cell types, including epithelial, endothelial, and mesenchymal cells.
Limitations and Considerations
While Dispase is a powerful tool, there are some limitations and considerations to keep in mind: Enzyme concentration: The effectiveness of Dispase can vary depending on the concentration used. It is essential to optimize the enzyme concentration for each specific tissue type.
Incubation time: Over-digestion can lead to cell damage and loss of viability. Therefore, it is crucial to monitor the incubation time carefully.
Batch variability: Different batches of Dispase may have varying enzyme activities, which can affect experimental consistency. It is advisable to test each batch before use.
Conclusion
Dispase is an essential tool in histology for tissue dissociation and cell isolation. Its ability to gently dissociate tissues while maintaining cell viability and functionality makes it invaluable for various applications, including immunohistochemistry and organoid culture. However, careful optimization of enzyme concentration and incubation time is necessary to achieve the best results.
