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Changes in the proteomic profile of adipose tissue-derived mesenchymal stem cells during passages

机译:传代过程中脂肪组织间充质干细胞的蛋白质组学变化

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Background Human mesenchymal stem cells (hMSC) have recently raised the attention because of their therapeutic potential in the novel context of regenerative medicine. However, the safety of these new and promising cellular products should be carefully defined before they can be used in the clinical setting, as. The protein expression profile of these cells might reveal potential hazards associated with senescence and tumoral transformation which may occur during culture. Proteomic is a valuable tool for hMSC characterization and identification of possible changes during expansion. Results We used Surface Enhanced Laser Desorption/Ionization-Time Of Flight-Mass Spectrometry (SELDI-ToF-MS) to evaluate the presence of stable molecular markers in adipose tissue-derived mesenchymal stem cells (AD-MSC) produced under conditions of good manufacturing practices (GMP). Proteomic patterns of cells prepared were consistent, with 4 up-regulated peaks (mass-to-charge ratio (m/z) 8950, 10087, 10345, and 13058) through subculture steps (P0-P7) with similar trend in three donors. Among the differentially expressed proteins found in the cytoplasmic and nuclear fractions, a cytoplasmic 10.1 kDa protein was upregulated during culture passages and was identified as S100A6 (Calcyclin). Conclusions This study suggests for the first time that common variation could occur in AD-MSC from different donors, with the identification of S100A6, a protein prevalently related to cell proliferation and cell culture condition. These results support the hypothesis of common proteomic changes during MSCs expansion and could give important insight in the knowledge of molecular mechanisms intervening during MSC expansion.
机译:背景技术人间充质干细胞(hMSC)由于在再生医学的新领域具有治疗潜力,最近引起了人们的关注。但是,在将这些新的有前途的细胞产品用于临床之前,应仔细定义其安全性。这些细胞的蛋白质表达特征可能揭示了与衰老和肿瘤转化相关的潜在危害,这些危害可能在培养过程中发生。蛋白质组学是用于hMSC表征和鉴定扩增过程中可能变化的有价值的工具。结果我们使用表面增强激光解吸/电离飞行时间质谱(SELDI-ToF-MS)来评估在良好制造条件下生产的脂肪组织间充质干细胞(AD-MSC)中稳定分子标记的存在实践(GMP)。制备的细胞的蛋白质组学模式是一致的,通过亚培养步骤(P0-P7)具有4个上调的峰(质荷比(m / z)8950、10087、10345和13058),三个供体的趋势相似。在细胞质和细胞核组分中发现的差异表达蛋白中,细胞质10.1 kDa蛋白在培养传代过程中被上调,被鉴定为S100A6(Calcyclin)。结论这项研究首次表明,通过鉴定S100A6(一种与细胞增殖和细胞培养条件普遍相关的蛋白),不同供体的AD-MSC可能发生共同变异。这些结果支持了MSC扩增过程中常见蛋白质组学变化的假说,并可能为了解MSC扩增过程中介入的分子机制提供重要见解。

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