In vivo culturing of a bilayered dermal substitute with adipo-stromal cells.
BACKGROUND
Skin grafting is an important procedure to cover skin defects. Recently, cultured epidermal sheets and bilayered cultured skin have been used clinically, but they lack subcutaneous tissue. The objective of this study was to produce a bilayered dermal substitute with adipose tissue simultaneously in vivo.


METHODS
We disseminated adipo-stromal cells on one side of a collagen sponge at a density of 1,0 x 10(5)cells/cm(2) and incubated overnight. Then, we turned over the sponge and disseminated dermal fibroblasts and keratinocytes at a density of 1,0 x 10(6)cells/cm(2) on the other side of the sponge. Finally, we cultured this for 1 wk and implanted it on the backs of severe combined immunodeficiency mice with or without basic FGF.


RESULTS
Six weeks after implantation, specimens were harvested. Macroscopically, the formed tissue in the bFGF-administered group was thick, and the epidermal component, the dermal component, and adipose tissue were formed in the cross section. The thickness of newly formed tissue in bFGF-administered group was significantly greater than that in the group without bFGF administration. The area of the newly formed capillaries in the bFGF-administered group was significantly larger than that in the group without bFGF administration.


CONCLUSIONS
We could produce a thick composite tissue in vivo, combining three kinds of human cells, collagen scaffold, and bFGF. This composite graft was thicker than the bilayered dermal substitute and could be a substitute for a skin flap.
Morimoto
Naoki
N
Department of Plastic and Reconstructive surgery, Graduate School of Medicine, Kyoto University, Kyoto, Japan. mnaoki22@kuhp.kyoto-u.ac.jp
Takemoto
Satoru
S
Kawazoe
Takeshi
T
Suzuki
Shigehiko
S
Tomihata
Kenji
K
Taira
Tsuguyoshi
T
D000273:Adipose Tissue / Q000166:cytology*; D000818:Animals; D002478:Cells, Cultured; D020405:Dermis / Q000166:cytology*; D005347:Fibroblasts / Q000502:physiology; D006801:Humans; D015603:Keratinocytes / Q000502:physiology; D051379:Mice; D016038:Skin Transplantation; D017768:Skin, Artificial; D017154:Stromal Cells / Q000502:physiology*