Methods:Adipose tissue-derived stem cells isolated from human liposuction aspirates using collagenase dispersion and used a tissue engineering skin system with human skin cells in a three-dimensional collagen gel matrix culture. We prepared human tissue engineering skin made of a collagen type I matrix of cattle skin populated either with human dermal fibroblasts or adipose tissue-derived stem cells (ADSCs), on top of which keratinocytes were seeded and subsequently grown at the air¨Cliquid interface. We observed ADSC- keratinocyte and ADSC-dermal fibroblasts interactions with histological method and morphometric analysis.
Results: In this reconstructed skin culture system, When keratinocytes were seeded on the ADSC and fibroblasts (cell number ratio,1:1) mixed layer, they proliferated extensively and formed a thick epidermal layer with a well-differentiated structure. When keratinocytes were seeded directly on ADSC layer without dermal fibroblasts, they were not well-differetiated than that seeded on the layer only with dermal fibroblasts. Conversely, ADSC promoted the proliferation of dermal fibroblasts. These effects of ADSC were presumed to be the multipotential of it and mediated by cytokines derived from ADSC or the differentiated cells.
Conclusions: The tissue engineering skin with different cell types can be used to observe cell-cell interactions and the mechanism of wound healing. Dermal fibroblast and ADSC may promote each self , dermal fibroblast may promote ADSC to differentiate into fat cell and endothelial cell. These cells secreted some cytokines to promoted the proliferation and differentiation of keratinocytes.