Reconstruction of three-dimensional human skin model composed of dendritic cells, keratinocytes and fibroblasts utilizing a handy scaffold of collagen vitrigel membrane.

We previously we attempted to make a three-dimensional human skin model consisting of three different cells, dendritic cells, keratinocytes and fibroblasts (KDF-Skin) to evaluate immunoreactions in human skin; however, this model had various problems; for example (1) the incubation period for the construction of this model is long (about three weeks); (2) to construct the collagen gel, high amounts of fibroblasts are needed; and (3) the horny layer of keratinocytes in this skin model is thinner than that of keratinocytes in real human skin. In order to overcome these problems, a new three-dimensional human skin model utilizing a handy scaffold of collagen vitrigel membrane (VG-KDF-Skin) was constructed. As a result, after 14 days incubation, the epidermis layer of normal human keratinocytes was thicker than the keratinocyte layer of KDF-Skin. The incubation period for VG-KDF-Skin construction was 7 days shorter than that of KDF-Skin, and the number of fibroblasts needed to seed VG-KDF-Skin was four times fewer than that of KDF-Skin. After the application of sensitizers such as DNCB, VG-KDF-Skin induced the expression of CD86 and cytokine release. These results suggest that the new three-dimensional human skin model consisting of dendritic cells, keratinocytes, fibroblasts and collagen vitrigel membrane was more useful for alternative animal testing than the KDF-Skin model.