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.
Uchino
T
T
National Institute of Health Sciences, Division of Environmental Chemistry, 1-18-1 Kamiyoga, Setaga-ku, Tokyo 158-8501, Japan. uchino@nihs.go.jp
Takezawa
T
T
Ikarashi
Y
Y
D000826:Animal Testing Alternatives; D051940:B7-2 Antigen / Q000378:metabolism; D018929:Cell Culture Techniques / Q000379:methods; D002478:Cells, Cultured; D018920:Coculture Techniques / Q000379:methods; D024042:Collagen Type I / Q000737:chemistry; D016207:Cytokines / Q000378:metabolism; D003713:Dendritic Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D005782:Gels; D006801:Humans; D007509:Irritants / Q000633:toxicity; D015603:Keratinocytes / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D008567:Membranes, Artificial; D008954:Models, Biological; D017768:Skin, Artificial; D013997:Time Factors; D054457:Tissue Scaffolds
D051940:B7-2 Antigen; D024042:Collagen Type I; D016207:Cytokines; D005782:Gels; D007509:Irritants; D008567:Membranes, Artificial