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Biomarkers Used to Detect Genetic Damage in Tissue-Engineered Skin

Published

Author(s)

C D. O'Connell, Peter E. Barker, M A. Marino, P McAndrew, Donald H. Atha, Pawel Jaruga, M Birincioglu, M. Dizdaroglu, H Rodriguez

Abstract

The goal of the research was to help identify cellular biomarkers and assess measurement technologies that will help manufacturers determine how cells react to inflammatory stimuli from different polymer matrixes and assure that tissue-engineered medical products are free of genetic changes that could occur during the manufacturing phase, storage or shipment of the product. In this study, tissue-engineered skin (TestSkin II ) was obtained, separated into its two cellular layers (epidermis and dermis) and DNA was extracted. The first biomarker tested consisted of screening for DNA point mutations in the p53 gene, the most commonly mutated gene in skin cancer. It was shown that tissue-engineered skin did not contain mutations in this gene at the level of sensitivity of SSCP-capillary electrophoresis and Denaturing High Performance Liquid Chromatography. Results were compared to control cells (neonatal fibroblasts and neonatal keratinocytes) and fibroblasts that were obtained from a 55 year-old and 96 year-old human donor. The second set of biomarkers tested looked at the loss of the Y-chromosome. Using Fluorescent In Situ Hybridization technology, no detectable loss of Y-chromosome was found in the tissue-engineered skin, neonatal and 55 year-old donor control cells. Y-chromosome loss was found in the fibroblasts from the 96 year-old donor. Biomarkers such as p53 mutations and chromosome loss can provide the basis for an international reference standard of cellular biomarkers that can aid in the development and safety of tissue engineered medical products.
Proceedings Title
Tissue Engineering, Stem Cells, and Gene Therapies
Volume
534
Conference Dates
September 19-22, 2002
Conference Location
Antalya, TR
Conference Title
9th International Symposium on Biomedical Science and Technology

Keywords

P53, tissue engineering, Y-chromosome

Citation

O'Connell, C. , Barker, P. , Marino, M. , McAndrew, P. , Atha, D. , Jaruga, P. , Birincioglu, M. , Dizdaroglu, M. and Rodriguez, H. (2003), Biomarkers Used to Detect Genetic Damage in Tissue-Engineered Skin, Tissue Engineering, Stem Cells, and Gene Therapies, Antalya, TR (Accessed April 19, 2024)
Created December 31, 2002, Updated October 12, 2021