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One-Step Preservation and Decalcification of Bony Tissue for Molecular Profiling

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Molecular Profiling

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1606))

Abstract

Bone metastasis from primary cancer sites creates diagnostic and therapeutic challenges. Calcified bone is difficult to biopsy due to tissue hardness and patient discomfort, thus limiting the frequency and availability of bone/bone marrow biopsy material for molecular profiling. In addition, bony tissue must be demineralized (decalcified) prior to histomorphologic analysis. Decalcification processes rely on three main principles: (a) solubility of calcium salts in an acid, such as formic or nitric acid; (b) calcium chelation with ethylenediaminetetraacetic acid (EDTA); or (c) ion-exchange resins in a weak acid. A major roadblock in molecular profiling of bony tissue has been the lack of a suitable demineralization process that preserves histomorphology of calcified and soft tissue elements while also preserving phosphoproteins and nucleic acids. In this chapter, we describe general issues relevant to specimen collection and preservation of osseous tissue for molecular profiling. We provide two protocols: (a) one-step preservation of tissue histomorphology and proteins and posttranslational modifications, with simultaneous decalcification of bony tissue, and (b) ethanol-based tissue processing for TheraLin-fixed bony tissue.

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Acknowledgments

We express our deepest gratitude to Antonella Chiechi, Ph.D., for facilitating experiments for evaluating fixation and decalcification methods with human bone specimens. This work was supported in part by the National Institutes of Health Innovative Molecular Analysis Technologies (IMAT) program through a grant to L. Liotta and V. Espina (1R33CA157403-01).

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Authors and Affiliations

  1. Center for Applied Proteomics and Molecular Medicine, George Mason University, 10920 George Mason Circle, MS1A9, Manassas, VA, 20110, USA

    Claudius Mueller, Michael G. Harpole & Virginia Espina

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  1. Claudius Mueller
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  2. Michael G. Harpole
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  3. Virginia Espina
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Correspondence to Claudius Mueller .

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  1. Center for Applied Proteomics and Molecular Medicine, George Mason University, Manassas, Virginia, USA

    Virginia Espina

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Mueller, C., Harpole, M.G., Espina, V. (2017). One-Step Preservation and Decalcification of Bony Tissue for Molecular Profiling. In: Espina, V. (eds) Molecular Profiling. Methods in Molecular Biology, vol 1606. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6990-6_6

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  • DOI: https://doi.org/10.1007/978-1-4939-6990-6_6

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6989-0

  • Online ISBN: 978-1-4939-6990-6

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