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. 2009 Mar;30(8):1587-95.
doi: 10.1016/j.biomaterials.2008.12.009. Epub 2008 Dec 23.

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

Wonhye Lee  1 , Jason Cushing Debasitis, Vivian Kim Lee, Jong-Hwan Lee, Krisztina Fischer, Karl Edminster, Je-Kyun Park, Seung-Schik Yoo
Affiliations

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

Wonhye Lee et al. Biomaterials. 2009 Mar.

Abstract

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 on 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 on a planar tissue culture dish, resulting in two distinct cell layers of inner fibroblasts and outer keratinocytes. In order to demonstrate the ability to print and culture multi-layered cell-hydrogel composites on a non-planar surface for potential applications including skin wound repair, the technique was tested on 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.

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