2D-DIGE Proteomic Analysis of Mesenchymal Stem Cell Cultured on the Elasticity-tunable Hydrogels
2D-DIGE Proteomic Analysis of Mesenchymal Stem Cell Cultured on the Elasticity-tunable Hydrogels
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DOI：
10.1247/csf.12012
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发表时间：
2012-01-01
影响因子：
1.5
通讯作者：
Kidoaki, Satoru
中科院分区：
文献类型：
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作者：
Kuboki, Thasaneeya;Kantawong, Fahsai;Kidoaki, Satoru
本研究的重点是间充质干细胞（MSCs）对基质弹性的机械转导反应。通过使用具有可调刚度的光固化凝胶，利用二维凝胶内差分分析（2D-DIGE）对组织培养塑料、软（3 kPa）和硬（52 kPa）基质上培养的MSCs的蛋白质组学图谱进行了解码。与免疫荧光相关的DIGE数据表明，细胞骨架蛋白的丰度和组织变化，以及重要信号相关蛋白、应激反应蛋白和胶原蛋白合成相关蛋白的差异调节。在凝胶上培养的细胞中，肌动蛋白（actin）、微管蛋白（tubulin）和波形蛋白（vimentin）等主要CSK蛋白的表达发生了显著变化。与刚性组织培养板相比，可以在凝胶样品上观察到α -微管蛋白和β -肌动蛋白的显著下调。在硬凝胶培养的MSCs中，波形蛋白的表达丰度最高。这些结果表明，基质刚度显著影响MSCs细胞骨架蛋白的表达平衡，并对细胞张拉完整性有一定影响。
The present study focuses on mechanotransduction in mesenchymal stem cells (MSCs) in response to matrix elasticity. By using photocurable gelatinous gels with tunable stiffness, proteomic profiles of MSCs cultured on tissue culture plastic, soft (3 kPa) and stiff (52 kPa) matrices were deciphered using 2-dimensional differential in-gel analysis (2D-DIGE). The DIGE data, tied to immunofluorescence, indicated abundance and organization changes in the cytoskeletonal proteins as well as differential regulation of important signalingrelated proteins, stress-responsing proteins and also proteins involved in collagen synthesis. The major CSK proteins including actin, tubulin and vimentin of the cells cultured on the gels were remarkably changed their expressions. Significant down-regulation of alpha-tubulin and beta-actin can be observed on gel samples in comparison to the rigid tissue culture plates. The expression abundance of vimentin appeared to be highest in the MSCs cultured on hard gels. These results suggested that the substrate stiffness significantly affects expression balances in cytoskeletal proteins of MSCs with some implications to cellular tensegrity.