Preparation of human epidermal tissue for functional immune studies.
A trypsinization technique was optimized for preparation of human epidermal single-cell suspensions from suction blisters. The conditions which provided the highest epidermal cell yield and viability was Sigma trypsin 0.25% (Type II, crude; cat. no. T8128). The incubation time should be 45 min at 37 degrees C with continuous shaking of the epidermal tissue. The use of suction blisters was evaluated for in vivo determination of epidermal thymocyte activating factor (ETAF/Il-1). Trypsinization of epidermis diminished ETAF/Il-1 activity. Dialysis of epidermal homogenates improves determination of ETAF/Il-1-like activity due to removal of low-weight inhibitors. Determination of ETAF/Il-1-like activity in homogenates of suction blisters constitutes a reliable model with acceptable reproducibility.
Larsen
C G
CG
Department of Dermatology, Marselisborg Hospital, University of Aarhus, Denmark.
Larsen
F G
FG
Thestrup-Pedersen
K
K
D002469:Cell Separation / Q000379:methods*; D000078404:Epidermal Cells; D004817:Epidermis / Q000032:analysis; D006801:Humans; D007375:Interleukin-1 / Q000032:analysis*; D013048:Specimen Handling; D014357:Trypsin
D007375:Interleukin-1; D014357:Trypsin