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工程化软骨的蛋白质组学分析

Proteomic Analysis of Engineered Cartilage.

作者信息

Pu Xinzhu, Oxford Julia Thom

机构信息

Department of Biological Sciences, Biomolecular Research Center, Boise State University, Boise, ID, USA.

Department of Biological Sciences, Biomolecular Research Center, Boise State University, 1910 University Drive, Mail Stop 1511, Boise, ID, USA.

出版信息

Methods Mol Biol. 2015;1340:263-78. doi: 10.1007/978-1-4939-2938-2_19.

Abstract

Tissue engineering holds promise for the treatment of damaged and diseased tissues, especially for those tissues that do not undergo repair and regeneration readily in situ. Many techniques are available for cell and tissue culturing and differentiation of chondrocytes using a variety of cell types, differentiation methods, and scaffolds. In each case, it is critical to demonstrate the cellular phenotype and tissue composition, with particular attention to the extracellular matrix molecules that play a structural role and that contribute to the mechanical properties of the resulting tissue construct. Mass spectrometry provides an ideal analytical method with which to characterize the full spectrum of proteins produced by tissue-engineered cartilage. Using normal cartilage tissue as a standard, tissue-engineered cartilage can be optimized according to the entire proteome. Proteomic analysis is a complementary approach to biochemical, immunohistochemical, and mechanical testing of cartilage constructs. Proteomics is applicable as an analysis approach to most cartilage constructs generated from a variety of cellular sources including primary chondrocytes, mesenchymal stem cells from bone marrow, adipose tissue, induced pluripotent stem cells, and embryonic stem cells. Additionally, proteomics can be used to optimize novel scaffolds and bioreactor applications, yielding cartilage tissue with the proteomic profile of natural cartilage.

摘要

组织工程有望用于治疗受损和患病组织,特别是那些在原位不易进行修复和再生的组织。利用多种细胞类型、分化方法和支架,有许多技术可用于软骨细胞的细胞和组织培养及分化。在每种情况下,证明细胞表型和组织组成至关重要,尤其要关注那些起结构作用并有助于所形成组织构建体力学性能的细胞外基质分子。质谱分析提供了一种理想的分析方法,可用于表征组织工程软骨产生的蛋白质全谱。以正常软骨组织为标准,可根据整个蛋白质组对组织工程软骨进行优化。蛋白质组学分析是对软骨构建体进行生化、免疫组织化学和力学测试的一种补充方法。蛋白质组学适用于分析由多种细胞来源产生的大多数软骨构建体,这些细胞来源包括原代软骨细胞、骨髓间充质干细胞、脂肪组织、诱导多能干细胞和胚胎干细胞。此外,蛋白质组学可用于优化新型支架和生物反应器应用,从而产生具有天然软骨蛋白质组特征的软骨组织。

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