Dendritic cells (DC) are crucial for the induction of immune responses and populate various tissues to fulfil their special role. The skin harbours different DC subsets, the Langerhans cells (LC) in the epidermis and the dermal DC in the dermis. The investigation of skin DC is cumbersome since these cells are rare in the skin. As a consequence, it is laborious to receive enough cells from the tissue for experiments. Several approaches have been developed to isolate skin DC based on either enzymatic digestion of the tissue or skin explant culture. Immature LC can be obtained by trypsinization of epidermis, cultured in vitro and be highly enriched with gradient centrifugation and magnetic bead sorting. Mature skin DC can be easily received from skin explant culture. For this purpose skin pieces are cultured for several days and migratory DC emigrate from epidermis and dermis. Both techniques are described for human and mouse skin in the following chapter of the book.