Gene Expression Patterns Related to Osteogenic Differentiation of Bone Marrow-Derived Mesenchymal Stem Cells During Ex Vivo Expansion
Gene Expression Patterns Related to Osteogenic Differentiation of Bone Marrow-Derived Mesenchymal Stem Cells During Ex Vivo Expansion
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DOI：
10.1089/ten.tec.2009.0405
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发表时间：
2010-06-01
影响因子：
3
通讯作者：
Ciapetti, Gabriela
中科院分区：
文献类型：
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作者：
Granchi, Donatella;Ochoa, Gorka;Ciapetti, Gabriela
骨髓通常被用作成人多潜能间充质干细胞(MSCs)的来源，MSCs的定义是其在体外分化为多个谱系的能力。在细胞治疗和组织工程应用方面，体外扩增的MSCs目前都被评估为修复受损骨骼组织功能的一种策略。本研究的目的是明确MSCs在体外扩增过程中向成熟成骨细胞分化的基因表达模式，并探索用形态、细胞化学和生化分析难以评估的各种细胞功能。从6名接受全髋关节置换术的患者的骨髓样本中获取细胞培养，并使用Affymetrix HG-U133A Plus 2.0阵列(Affymetrix(R)，Santa Clara，CA)在发生特定事件时进行大规模转录组分析，包括MSC表面标记的出现、克隆的形成和矿物质结节的沉积。我们的注意力集中在213个差异表达的基因上，其中一些属于众所周知的通路，一些具有一个或多个与骨细胞生物学相关的基因本体论注释，包括血管生成、骨相关基因、细胞通讯、发育和形态发生、转化生长因子-β信号和Wnt信号。已发现29个基因，其在骨细胞病理生理学中的作用尚未被描述。总之，定义了表征体外扩增的MSCs成骨分化过程的早期、中期和晚期的基因表达模式。这些信号是监测成骨过程和分析支架上培养的MSCs的广泛功能的有用工具，特别是当构建的结构是为了释放生长因子或其他促进骨再生的信号时。
Bone marrow is commonly used as a source of adult multipotent mesenchymal stem cells (MSCs), defined for their ability to differentiate in vitro into multiple lineages. The ex vivo-expanded MSCs are currently being evaluated as a strategy for the restoration of function in damaged skeletal tissue, both in cell therapy and tissue engineering applications. The aim of this study was to define gene expression patterns underlying the differentiation of MSCs into mature osteoblasts during the expansion in vitro, and to explore a variety of cell functions that cannot be easily evaluated using morphological, cytochemical, and biochemical assays. Cell cultures were obtained from bone marrow samples of six individuals undergoing total hip replacement, and a large-scale transcriptome analysis, using Affymetrix HG-U133A Plus 2.0 array (Affymetrix (R), Santa Clara, CA), was performed at the occurrence of specific events, including the appearance of MSC surface markers, formation of colonies, and deposition of mineral nodules. We focused our attention on 213 differentially upregulated genes, some belonging to well-known pathways and some having one or more Gene Ontology annotations related to bone cell biology, including angiogenesis, bone-related genes, cell communication, development and morphogenesis, transforming growth factor-beta signaling, and Wnt signaling. Twenty-nine genes, whose role in bone cell pathophysiology has not been described yet, were found. In conclusion, gene expression patterns that characterize the early, intermediate, and late phases of the osteogenic differentiation process of ex vivo-expanded MSCs were defined. These signatures represent a useful tool to monitor the osteogenic process, and to analyze a broad spectrum of functions of MSCs cultured on scaffolds, especially when the constructs are conceived for releasing growth factors or other signals to promote bone regeneration.