One-Step Preservation and Decalcification of Bony Tissue for Molecular Profiling.
One-Step Preservation and Decalcification of Bony Tissue for Molecular Profiling.
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用于分子分析的骨组织的一步保存和脱钙。
DOI：
10.1007/978-1-4939-6990-6_6
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发表时间：
2017
期刊：
影响因子：
--
通讯作者：
Espina,Virginia
中科院分区：
文献类型：
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作者：
Mueller,Claudius;Harpole,MichaelG;Espina,Virginia
原发癌部位的骨转移给诊断和治疗带来了挑战。由于组织硬度和患者不适，钙化骨很难进行活检，从而限制了骨/骨髓活检材料用于分子轮廓分析的频率和可用性。此外，在进行组织形态分析之前，骨组织必须脱矿(脱钙)。脱钙过程依赖于三个主要原则：(A)钙盐在甲酸或硝酸等酸中的溶解度；(B)钙与乙二胺四乙酸(EDTA)的络合作用；或(C)离子交换树脂在弱酸中的作用。骨组织分子图谱的一个主要障碍是缺乏合适的脱矿过程，该过程既能保留钙化和软组织成分的组织形态，又能保存磷蛋白和核酸。在这一章中，我们描述了与收集标本和保存骨组织相关的一般问题，以用于分子图谱。我们提供了两种方案：(A)一步保存组织的组织形态和蛋白质以及翻译后修改，同时对骨组织进行脱钙；(B)对TheraLin固定的骨组织进行基于乙醇的组织处理。
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.