A co-cultured skin model based on cell support membranes

Niann Tzyy Dai, Ming Kung Yeh, Demeral David Liu, E. F. Adams, Chiao Hsi Chiang, Chung Yang Yen, Chien Ming Shih, Huey Kang Sytwu, Tim Mo Chen, Hsian-Jenn Wang, M. R. Williamson, A. G.A. Coombes

Research output: Contribution to journalArticle

27 Citations (Scopus)

Abstract

Tissue engineering of skin based on collagen:PCL biocomposites using a designed co-culture system is reported. The collagen:PCL biocomposites having collagen:PCL (w/w) ratios of 1:4, 1:8, and 1:20 have been proven to be biocompatible materials to support both adult normal human epidermal Keratinocyte (NHEK) and mouse 3T3 fibroblast growth in cell culture, respectively, by Dai, Coombes, et al. in 2004. Films of collagen:PCL biocomposites were prepared using non-crosslinking method by impregnation of lyophilized collagen mats with PCL/dichloromethane solutions followed by solvent evaporation. To mimic the dermal/epidermal structure of skin, the 1:20 collagen:PCL biocomposites were selected for a feasibility study of a designed co-culture technique that would subsequently be used for preparing fibroblast/biocomposite/keratinocyte skin models. A 55.3% increase in cell number was measured in the designed co-culture system when fibroblasts were seeded on both sides of a biocomposite film compared with cell culture on one surface of the biocomposite in the feasibility study. The co-culture of human keratinocytes and 3T3 fibroblasts on each side of the membrane was therefore studied using the same co-culture system by growing keratinocytes on the top surface of membrane for 3 days and 3T3 fibroblasts underneath the membrane for 6 days. Scanning electron microscopy (SEM) and immunohistochemistry assay revealed good cell attachment and proliferation of both human keratinocytes and 3T3 fibroblasts with these two types of cells isolated well on each side of the membrane. Using a modified co-culture technique, a co-cultured skin model presenting a confluent epidermal sheet on one side of the biocomposite film and fibroblasts populated on the other side of the film was developed successfully in co-culture system for 28 days under investigations by SEM and immunohistochemistry assay. Thus, the design of a co-culture system based on 1:20 (w/w) collagen:PCL biocomposite membranes for preparation of a bi-layered skin model with differentiated epidermal sheet was proven in principle. The approach to skin modeling reported here may find application in tissue engineering and screening of new pharmaceuticals.

Original languageEnglish
Pages (from-to)905-908
Number of pages4
JournalBiochemical and Biophysical Research Communications
Volume329
Issue number3
DOIs
Publication statusPublished - Apr 15 2005
Externally publishedYes

Fingerprint

Fibroblasts
Coculture Techniques
Skin
Collagen
Cell Membrane
Membranes
Keratinocytes
Cell culture
Culture Techniques
Tissue engineering
Feasibility Studies
Tissue Engineering
Assays
Electron Scanning Microscopy
Cell Culture Techniques
Immunohistochemistry
Scanning electron microscopy
Methylene Chloride
Biocompatible Materials
Impregnation

Keywords

  • Biocomposite
  • Co-culture
  • Collagen
  • Fibroblast
  • Keratinocyte
  • Polycaprolactone

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Dai, N. T., Yeh, M. K., Liu, D. D., Adams, E. F., Chiang, C. H., Yen, C. Y., ... Coombes, A. G. A. (2005). A co-cultured skin model based on cell support membranes. Biochemical and Biophysical Research Communications, 329(3), 905-908. https://doi.org/10.1016/j.bbrc.2005.02.059

A co-cultured skin model based on cell support membranes. / Dai, Niann Tzyy; Yeh, Ming Kung; Liu, Demeral David; Adams, E. F.; Chiang, Chiao Hsi; Yen, Chung Yang; Shih, Chien Ming; Sytwu, Huey Kang; Chen, Tim Mo; Wang, Hsian-Jenn; Williamson, M. R.; Coombes, A. G.A.

In: Biochemical and Biophysical Research Communications, Vol. 329, No. 3, 15.04.2005, p. 905-908.

Research output: Contribution to journalArticle

Dai, NT, Yeh, MK, Liu, DD, Adams, EF, Chiang, CH, Yen, CY, Shih, CM, Sytwu, HK, Chen, TM, Wang, H-J, Williamson, MR & Coombes, AGA 2005, 'A co-cultured skin model based on cell support membranes', Biochemical and Biophysical Research Communications, vol. 329, no. 3, pp. 905-908. https://doi.org/10.1016/j.bbrc.2005.02.059
Dai, Niann Tzyy ; Yeh, Ming Kung ; Liu, Demeral David ; Adams, E. F. ; Chiang, Chiao Hsi ; Yen, Chung Yang ; Shih, Chien Ming ; Sytwu, Huey Kang ; Chen, Tim Mo ; Wang, Hsian-Jenn ; Williamson, M. R. ; Coombes, A. G.A. / A co-cultured skin model based on cell support membranes. In: Biochemical and Biophysical Research Communications. 2005 ; Vol. 329, No. 3. pp. 905-908.
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AU - Shih, Chien Ming

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