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, Sytwua, Huey-Kang, Chen, Tim-Mo, Wang, Hsian-Jenn, Williamson, M. R. and Coombes, A. G. A. (2005) A Co-cultured Skin Model Based on Cell Support Membranes. Biochemical and Biophysical Research Communication, 329 3: 905-908.


Author Dai, Niann-Tzyy
Yeh, Ming-Kung
Liu, Demeral David
Adams, E. F.
Chiang, Chiao-Hsi
Yen, Chung-Yang
Shih, Chien-Ming
Sytwua, Huey-Kang
Chen, Tim-Mo
Wang, Hsian-Jenn
Williamson, M. R.
Coombes, A. G. A.
Title A Co-cultured Skin Model Based on Cell Support Membranes
Journal name Biochemical and Biophysical Research Communication   Check publisher's open access policy
ISSN 0006-291X
Publication date 2005-04-15
Sub-type Article (original research)
DOI 10.1016/j.bbrc.2005.02.059
Volume 329
Issue 3
Start page 905
End page 908
Total pages 4
Place of publication New York
Publisher Academic Press
Language eng
Subject 111504 Pharmaceutical Sciences
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.
Keyword Biophysics
Biochemistry
Co-culture
Collagen
Polycaprolactone
Biocomposite
Keratinocyte
Fibroblast
Q-Index Code C1

Document type: Journal Article
Sub-type: Article (original research)
Collections: Excellence in Research Australia (ERA) - Collection
School of Pharmacy Publications
 
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