Separation of the epidermal sheet by dispase

Area of Science:

• Biotechnology
• Dermatology
• Cell Biology

Background:

• Dispase, a neutral protease from Bacillus polymyxa, is utilized in tissue separation.
• Efficient methods for isolating viable human epidermal cells are crucial for research and regenerative medicine.

Purpose of the Study:

• To evaluate the efficacy of dispase in separating human epidermis from the dermis.
• To assess the structural integrity and viability of epidermal cells post-separation.
• To establish a method for culturing keratinocytes from dispase-isolated epidermis.

Main Methods:

• Human skin explants were treated with dispase (500 and 1000 U/ml) for 24 hours.
• Epidermal sheets were analyzed using light and electron microscopy.
• Separated epidermal sheets were incubated with trypsin to obtain cell suspensions.
• Cell viability was assessed using the trypan blue dye exclusion test.
• Keratinocyte cultures were established from the cell suspensions.

Main Results:

• Dispase treatment enabled easy separation of the epidermis, retaining rete ridges.
• Electron microscopy revealed intact desmosomes and cells with villi and cytoplasmic projections.
• Epidermal cell suspensions showed over 95% viability.
• Successful keratinocyte cultures were established from the isolated cells.

Conclusions:

• Dispase is an effective enzyme for separating human epidermis while preserving cellular morphology and viability.
• The method facilitates the isolation of a high-yield, viable epidermal cell suspension suitable for keratinocyte culture.
• This technique offers a promising approach for obtaining cells for dermatological research and therapeutic applications.