Collection of skin samples for Flow Cytometric analysis (FACS).
A skin sample corresponding to the patch application area was harvested
using an 8-mm disposal biopsy punch (KAI medical) and transferred into 1
mL of Liberase TM (Roche) prepared in basic medium (RPMI + PS + 55 µM
BME + 20 mM HEPES), then incubated for 2 hours at 37°C. The enzymatic
reaction was then stopped with 75 µL of EDTA (100 mM). Cells were
homogenized using the Medimachine tissue homogenizer (BD Bioscience) for
8 min.