STP1452: Molecular Biomarkers Used to Detect Cellular/Genetic Damage in Tissue-Engineered Skin

    O'Connell, C
    Staff Scientist, Staff Scientist, Staff Scientist, and Staff Scientist, National Institute of Standards and Technology, Gaithersburg, MD

    Barker, PE
    Staff Scientist, Staff Scientist, Staff Scientist, and Staff Scientist, National Institute of Standards and Technology, Gaithersburg, MD

    Marino, M
    Transgenomic, Inc., Gaithersburg, MD

    McAndrew, P
    Transgenomic, Inc., Gaithersburg, MD

    Atha, DH
    Staff Scientist, Staff Scientist, Staff Scientist, and Staff Scientist, National Institute of Standards and Technology, Gaithersburg, MD

    Jaruga, P
    Guest Researcher, University of Maryland, Baltimore County,

    Birincioglu, M
    Medical School, Inönü University, Malatya,

    Rodriguez, H
    Staff Scientist, Staff Scientist, Staff Scientist, and Staff Scientist, National Institute of Standards and Technology, Gaithersburg, MD

    Pages: 8    Published: Jan 2004


    Abstract

    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 exons 5 and 6 of the TP53 gene, the most commonly mutated gene in skin cancer. To ensure the accuracy of the results, two measurement technologies that incorporate internal calibration standards were used. 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 and neonatal control cells. Y-chromosome loss was found in the fibroblasts from the 96-year-old donor. Biomarkers such as TP53 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.

    Keywords:

    TP53, CE-SSCP, DHPLC, Y-chromosome, biomarkers


    Paper ID: STP11650S

    Committee/Subcommittee: F04.40

    DOI: 10.1520/STP11650S


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