Biotrack S,r,l, Parco Tecnologico Padano, Lodi, Italy.
Proteome Sci. 2012 Jul 24;10(1):46. doi: 10.1186/1477-5956-10-46.
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.
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).
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.
人类间充质干细胞(hMSC)因其在再生医学新背景下的治疗潜力而受到关注。然而,在这些新的有前途的细胞产品可用于临床环境之前,应仔细定义其安全性,因为这些细胞的蛋白质表达谱可能揭示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 hMSC 特征描述和鉴定细胞在扩增过程中可能发生的变化的有用工具。
我们使用表面增强激光解吸/电离-飞行时间质谱(SELDI-ToF-MS)来评估在良好生产规范(GMP)条件下生产的脂肪组织来源间充质干细胞(AD-MSC)中稳定分子标志物的存在。通过亚培养步骤(P0-P7)制备的细胞的蛋白质组学模式一致,有 4 个上调峰(质荷比(m/z)8950、10087、10345 和 13058),在三个供体中具有相似的趋势。在细胞质和核部分发现的差异表达蛋白中,一种在培养过程中上调的细胞质 10.1 kDa 蛋白被鉴定为 S100A6(钙调蛋白)。
这项研究首次表明,来自不同供体的 AD-MSC 可能会发生常见的变异,鉴定出 S100A6,这是一种与细胞增殖和细胞培养条件密切相关的蛋白。这些结果支持了在 MSC 扩增过程中存在常见蛋白质组学变化的假设,并为理解 MSC 扩增过程中涉及的分子机制提供了重要的见解。