The various types of cells exist in three-layer structure, i. e., epidermis, dermis and subcutaneous tissue of human skin. Keratinocytes, melanocytes, Langerhans cells and Merkel cells are in the epidermis, fibroblasts, mast cells, histiocyte, plasma cells and dermal dendrocytes in the dermis, and adipocytes in the subcutaneous tissue. The dermal fibroblasts are isolated by outgrowth derived from dermis tissue for primary culture and can be repeatedly passaged with serum-containing medium. The epidermal keratinocytes are obtained from trypsinization of fullthickness skin biopsy. The serum-containing medium with some growth factors and feeder layer of 3T3 cells, so called Greenʼs method, can offer an environment for the preferable proliferation of keratinocytes. The keratinocytes cultured with serum-free medium become differentiated under increased calcium concentration. The melanocytes are easily cultured in the commercial medium without serum. But the melanocytes cultured with keratinocyte by the Greenʼs method can only reconstruct an intercellular function. Cells obtained from human skin are useful tools in various cell therapies.
