Model for human skin reconstructed in vitro composed of associated dermis and epidermis.
OBJECTIVE
The technique of obtaining human skin with dermis and epidermis reconstructed from cells isolated from patients can enable autologous skin grafting on patients with few donor sites. It also enables in vitro trials on chemicals and drugs. The objective of this work was to demonstrate a method for obtaining human skin composed of associated dermis and epidermis, reconstructed in vitro.


METHODS
Experimental laboratory study, in the Skin Cell Culture Laboratory of Faculdade de Ciências Médicas, Universidade Estadual de Campinas.


METHODS
Cells from human fibroblast cultures are injected into bovine collagen type I matrix and kept immersed in specific culturing medium for fibroblasts. This enables human dermis reconstruction in vitro. On this, by culturing human keratinocytes and melanocytes, differentiated epidermis is formed, leading to the creation of human skin composed of associated dermis and epidermis, reconstructed in vitro.


RESULTS
We showed that human skin composed of associated dermis and epidermis can be successfully reconstructed in vitro. It is histologically formed in the same way as human skin in vivo. Collagen tissue can be identified in the dermis, with cells and extracellular matrix organized in parallel to multilayer epidermis.


CONCLUSIONS
It is possible to obtain completely differentiated human skin composed of associated dermis and epidermis, reconstructed in vitro, from injection of human fibroblasts into bovine collagen type I matrix and culturing of human keratinocytes and melanocytes on this matrix.
Souto
Luís Ricardo Martinhão
LR
Skin Cell Culture Laboratory, Faculdade de Ciências Médicas, Universidade Estadual de Campinas (Unicamp), Campinas, São Paulo, Brazil. lrmsouto@yahoo.com.br
Rehder
Jussara
J
Vassallo
José
J
Cintra
Maria Letícia
ML
Kraemer
Maria Helena Stangler
MH
Puzzi
Maria Beatriz
MB
D000818:Animals; D002417:Cattle; D024042:Collagen Type I; D020405:Dermis / Q000166:cytology*; D000078404:Epidermal Cells; D005109:Extracellular Matrix; D005347:Fibroblasts / Q000166:cytology*; D006801:Humans; D007150:Immunohistochemistry; D015603:Keratinocytes / Q000166:cytology; D008544:Melanocytes / Q000166:cytology; D023822:Tissue Engineering / Q000379:methods*
D024042:Collagen Type I