Comparison of the effect of a proteolytic enzyme dispase and a chelator for the preparation of epidermal sheet from nude mice.
Histochemical studies of epidermal Langerhans cells require well-preserved epidermal sheets. We studied the conditions for the preparation of the epidermal sheets from the skin of the ear, hind limb and trunk of nude mice. Two types of commercial dispase, and ethylenediaminetetraacetic acid (EDTA), were used at several concentrations and the effects were compared. Dermo-epidermal separation was evaluated by the preservation of ATPase activity of Langerhans cells in the epidermal sheet and by the ultrastructure of epidermal cells (Langerhans cells and keratinocytes) as well as epidermal-dermal junction. Good separation depended on the combination of the concentration of the reagent and site of the skin. The concentrations of both dispase and EDTA effective for separation of the epidermis in the nude mouse were lower than those in other mouse strains reported.
Fujita
Y
Y
Central Laboratory for Electron Microscopy, Nagoya University School of Medicine, Japan.
Kobayashi
M
M
Kobayashi
K
K
Hoshino
T
T
D000251:Adenosine Triphosphatases / Q000032:analysis; D000818:Animals; D002469:Cell Separation / Q000379:methods; D002614:Chelating Agents / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D004423:Ear; D004492:Edetic Acid / Q000494:pharmacology*; D004591:Electrophoresis, Polyacrylamide Gel; D010450:Endopeptidases / Q000494:pharmacology*; D000078404:Epidermal Cells; D004817:Epidermis / Q000187:drug effects* / Q000648:ultrastructure; D005260:Female; D006614:Hindlimb; D006651:Histocytochemistry; D015603:Keratinocytes / Q000166:cytology / Q000648:ultrastructure; D007801:Langerhans Cells / Q000187:drug effects / Q000201:enzymology / Q000648:ultrastructure; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D008854:Microscopy, Electron; D010447:Peptide Hydrolases / Q000494:pharmacology*; D013909:Thorax
D002614:Chelating Agents; D004492:Edetic Acid; D010450:Endopeptidases; D010447:Peptide Hydrolases; C023139:dispase; D000251:Adenosine Triphosphatases