Isolation of a viable eccrine sweat gland by dispase

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Abstract
Dispase was used to obtain viable eccrine sweat glands from human skin in an intact shape. The full thickness of human skin was soaked in a solution of dispase in Eagle's minimum essential medium at a concentration of 500 units/ml and kept in a refrigerator at 4°C for 24h. The epidermal sheet with its appendages could then be easily separated from the dermis by lifting the epidermis with fine forceps. Electronmicroscopic observation revealed that the eccrine sweat gland was completely separated from the dermis at the basement membrane zone. The isolated epidermal sheet was scarcely dissociated by mechanical agitation in the presence of Ca2+ and Mg2+ ions. The eccrine sweat gland was cut away from the epidermis by using microscissors under a stereomicroscope. A cell suspension of the isolated eccrine sweat glands was obtained after trypsinization. The cells remained more than 90% viable up to 48 h in the culture medium. The obtained viable eccrine sweat glands will be useful for the study of the biology of sweating.