Stoitzner Patrizia, Romani Nikolaus, McLellan Alexander D, Tripp Christoph H, Ebner Susanne
Department of Dermatology and Venereology, Innsbruck Medical University, Innsbruck, Austria.
Methods Mol Biol. 2010;595:235-48. doi: 10.1007/978-1-60761-421-0_16.
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.
树突状细胞(DC)对于免疫反应的诱导至关重要,并分布于各种组织以履行其特殊作用。皮肤中存在不同的DC亚群,即表皮中的朗格汉斯细胞(LC)和真皮中的真皮DC。由于这些细胞在皮肤中数量稀少,因此对皮肤DC的研究很麻烦。因此,从组织中获取足够的细胞用于实验很费力。已经开发了几种方法来基于组织的酶消化或皮肤外植体培养来分离皮肤DC。未成熟的LC可以通过表皮胰蛋白酶消化获得,在体外培养,并通过梯度离心和磁珠分选高度富集。成熟的皮肤DC可以很容易地从皮肤外植体培养中获得。为此,将皮肤片培养数天,迁移性DC从表皮和真皮中移出。本书的下一章将介绍针对人类和小鼠皮肤的这两种技术。