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

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

Research output: Contribution to journalArticle

269 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)1587-1595
Number of pages9
JournalBiomaterials
Volume30
Issue number8
DOIs
Publication statusPublished - 2009 Mar 1
Externally publishedYes

Fingerprint

Fibroblasts
Keratinocytes
Cell culture
Skin
Fabrication
Tissue culture
Hydrogel
Collagen
Hydrogels
Cells
Tissue
Sodium bicarbonate
Printing
Sodium Bicarbonate
Drug Design
Wounds and Injuries
Composite materials
Robotics
Polydimethylsiloxane
Grafts

Keywords

  • 3D freeform fabrication
  • Collagen hydrogel
  • Fibroblasts
  • Keratinocytes
  • Skin tissue regeneration
  • Tissue engineering

ASJC Scopus subject areas

  • Biomaterials
  • Bioengineering
  • Ceramics and Composites
  • Mechanics of Materials
  • Biophysics

Cite this

Multi-layered culture of human skin fibroblasts and keratinocytes through three-dimensional freeform fabrication. / Lee, Wonhye; Debasitis, Jason Cushing; Lee, Vivian Kim; Lee, Jong-Hwan; Fischer, Krisztina; Edminster, Karl; Park, Je Kyun; Yoo, Seung Schik.

In: Biomaterials, Vol. 30, No. 8, 01.03.2009, p. 1587-1595.

Research output: Contribution to journalArticle

Lee, Wonhye ; Debasitis, Jason Cushing ; Lee, Vivian Kim ; Lee, Jong-Hwan ; Fischer, Krisztina ; Edminster, Karl ; Park, Je Kyun ; Yoo, Seung Schik. / Multi-layered culture of human skin fibroblasts and keratinocytes through three-dimensional freeform fabrication. In: Biomaterials. 2009 ; Vol. 30, No. 8. pp. 1587-1595.
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