Transplantation of Composite Skin Containing Keratinocytes Cultured on a Fibroblast-conditioned Acellular Dermal Matrix.

To evaluate the role of fibroblasts in composite skin reconstructed in vitro, four different types of composite skin (A, B, C, and D) were prepared. Human keratinocytes were seeded onto the epidermal side of an acellular dermal matrix (ADM) in type A. Keratinocytes were seeded onto the epidermal side of an ADM and human fibroblasts onto the dermal side in type B. Both keratinocytes and fibroblasts were seeded onto the epidermal side in type C. Type D consisted of fibroblasts on both sides of the ADM and keratinocytes on the epidermal side. The adherence of keratinocytes to the ADM was observed. The composite skin was then transplanted onto full-thickness skin defect wounds in nude mice. Results showed that the adherence of keratinocytes to the ADM was improved when fibroblasts were pre-seeded onto the epidermal side of the ADM. The composite skin was able to close full-thickness skin defect wounds. The take rates were respectively 44.1 ± 7.8%, 47.3 ± 5.4%, 75.2 ± 8.8%, and 81.2 ± 8.1% for types A, B, C, and D. The take rates of types C and D were significantly higher than those of types A and D. There was no significant difference in take rate between types C and D. In conclusion, composite skin consisting of keratinocytes cultured on a fibroblast-conditioned ADM was a good skin substitute.