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首页> 外文期刊>Proteomics >Differential expression of cell surface proteins in human bone marrow mesenchymal stem cells cultured with or without basic fibroblast growth factor containing medium
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Differential expression of cell surface proteins in human bone marrow mesenchymal stem cells cultured with or without basic fibroblast growth factor containing medium

机译:含或不含碱性成纤维细胞生长因子培养基培养的人骨髓间充质干细胞中细胞表面蛋白的差异表达

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Mesenchymal stem cells (MSCs) are multipotent cells, which have the capability to differentiate into various mesenchymal tissues such as bone, cartilage, fat, tendon, muscle, and marrow stroma. However, they lose the capability of multi-lineage differentiation after several passages. It is known that basic fibroblast growth factor (bFGF) increases growth rate, differentiation potential, and morphological changes of MSCs in vitro. In this report, we have used 2-DE coupled to MS to identify differentially expressed proteins at the cell membrane level in MSCs growing in bFGF containing medium. The cell surface proteins isolated by the biotin-avidin affinity column were separated by 2-DE in triplicate experiments. A total of 15 differentially expressed proteins were identified by quadrupole-time of flight tandem MS. Nine of the proteins were upregulated and six proteins were downregulated in the MSCs cultured with bFGF containing medium. The expression level of three actin-related proteins, F-actin-capping protein subunit alpha-1, actin-related protein 2/3 complex subunit 2, and myosin regulatory light chain 2, was confirmed by Western blot analysis. The results indicate that the expression levels of F-actin-capping protein subunit alpha-1, actin-related protein 2/3 complex subunit 2, and myosin regulatory light chain 2 are important in bFGF-induced morphological change of MSCs.
机译:间充质干细胞(MSC)是多能细胞,具有分化为各种间充质组织的能力,例如骨骼,软骨,脂肪,肌腱,肌肉和骨髓基质。然而,它们在经过几代之后失去了多系分化的能力。众所周知,碱性成纤维细胞生长因子(bFGF)可提高MSC的生长速率,分化潜能和体外形态学变化。在本报告中,我们使用了2-DE偶联MS来鉴定在包含bFGF的培养基中生长的MSC中细胞膜水平的差异表达蛋白。通过生物素-亲和素亲和柱分离的细胞表面蛋白通过2-DE在三次重复实验中分离。通过四极杆飞行时间串联质谱仪鉴定出总共15种差异表达的蛋白质。在用含bFGF的培养基培养的MSC中,九种蛋白质被上调,六种蛋白质被下调。通过蛋白质印迹分析证实了三种肌动蛋白相关蛋白,F-肌动蛋白封闭蛋白亚基α-1,肌动蛋白相关蛋白2/3复合物亚基2和肌球蛋白调节轻链2的表达水平。结果表明,F-肌动蛋白封端蛋白亚基α-1,肌动蛋白相关蛋白2/3复杂亚基2和肌球蛋白调节轻链2的表达水平在bFGF诱导的MSC形态改变中很重要。

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