Multi-layered culture of human skin fibroblasts and keratinocytes through three-dimensional freeform fabrication

We present a method to create multi-layered engineered tissue composites consisting of human skin fibroblasts and keratinocytes which mimic skin layers. Three-dimensional (3D) freeform fabrication (FF) technique, based oil direct cell dispensing, was implemented using a robotic platform that prints collagen hydrogel precursor, fibroblasts and keratinocytes. A printed layer of cell-containing collagen was crosslinked by coating the layer with nebulized aqueous sodium bicarbonate. The process was repeated in layer-by-layer fashion oil a planar tissue Culture fish, resulting ill two distinct cell layers of inner fibroblasts and outer keratinocytes. it order to demonstrate the ability to print and Culture multi-layer cell-hydrogel composites oil a non-planar surface for potential applications including skin Wound repair, the technique Was tested oil a poly(dimethylsiloxane) (PDMS) mold with 3D surface Contours as a target Substrate. Highly viable proliferation of each cell layer was observed on both planar and non-planar surfaces. Our results Suggest that organotypic skin tissue Culture is feasible using on-demand cell printing technique with future potential application in creating skin grafts tailored for Wound shape or artificial tissue assay for disease modeling and drug testing. (C) 2008 Elsevier Ltd. All rights reserved.