Effects of hormone and growth factor on formation of fibroblast-aggregates for tissue-engineered skin

To accelerate wound healing after implantation of tissue-engineered skin products, higher density inoculation of fibroblasts would be important. Therefore, we attempted to develop a tissue-engineered skin with a higher density of fibroblasts by formation of cell aggregates combined with a biodegradable polymer. Our method consisted of rotationally shaking with nontreated dishes, declining fibroblast-material interactions, and augmenting cell–cell interaction. Under the culture condition, normal neonatal human dermal fibroblasts formed spheroidal aggregates. The formation of the fibroblast-aggregates was accelerated by the addition of insulin, dexamethasone, basic fibroblast growth factor, and ascorbic acid to basal medium, and by the control of shaking time. In fibroblast-aggregates formed under a basal medium, the level of gene expression related to wound healing was higher than that of a monolayer when one added the hormones and growth factors. Therefore, tissue-engineered skin products with aggregates and scaffolds would become an artificial skin with potentiality of higher wound healing.