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在体外扩增过程中,肾祖细胞比间充质干细胞具有更高的遗传稳定性和更低的氧化应激。

Renal Progenitor Cells Have Higher Genetic Stability and Lower Oxidative Stress than Mesenchymal Stem Cells during In Vitro Expansion.

机构信息

Integrated Nucleus of Morphology and Stem Cell Research (NUPCelt), Center for Agrarian Sciences, Federal University of Piauí, Brazil.

Federal Institute of Education, Science and Technology of Piauí, Brazil.

出版信息

Oxid Med Cell Longev. 2020 Jul 10;2020:6470574. doi: 10.1155/2020/6470574. eCollection 2020.

Abstract

In vitro senescence of multipotent cells has been commonly associated with DNA damage induced by oxidative stress. These changes may vary according to the sources of production and the studied lineages, which raises questions about the effect of growing time on genetic stability. This study is aimed at evaluating the evolution of genetic stability, viability, and oxidative stress of bone marrow mesenchymal stem cells (MSCBMsu) and renal progenitor cells of the renal cortex (RPCsu) of swine (Sus scrofa domesticus) in culture passages. P2, P5, and P9 were used for MSCBMsu and P1, P2, and P3 for RPCsu obtained by thawing. The experimental groups were submitted to MTT, apoptosis and necrosis assays, comet test, and reactive substance measurements of thiobarbituric acid (TBARS), nitrite, reduced glutathione (GSH), and catalase. The MTT test curve showed a mean viability of 1.14 ± 0.62 and 1.12 ± 0.54, respectively, for MSCBMsu and RPCsu. The percentages of MSCBMsu and RPCsu were presented, respectively, for apoptosis, an irregular and descending behavior, and necrosis, ascending and irregular. The DNA damage index showed higher intensity among the MSCBMsu in the P5 and P9 passages ( < 0.05). In the TBARS evaluation, there was variation among the lines of RPCsu and MSCBMsu, presenting the last most significant variations ( < 0.05). In the nitrite values, we identified only among the lines, in the passages P1 and P2, with the highest averages displayed by the MSCBMsu lineage ( < 0.05). The measurement of antioxidant system activity showed high standards, identifying differences only for GSH values, in the RPCsu lineage, in P3 ( < 0.05). This study suggests that the maintenance of cell culture in the long term induces lower regulation of oxidative stress, and RPCsu presents higher genetic stability and lower oxidative stress than MSCBMsu during in vitro expansion.

摘要

体外衰老的多能细胞通常与氧化应激诱导的 DNA 损伤有关。这些变化可能因生产来源和研究谱系的不同而有所不同,这就提出了关于生长时间对遗传稳定性影响的问题。本研究旨在评估猪骨髓间充质干细胞(MSCBMsu)和肾皮质肾祖细胞(RPCsu)在培养传代过程中的遗传稳定性、活力和氧化应激的演变。P2、P5 和 P9 用于解冻获得的 MSCBMsu,P1、P2 和 P3 用于 RPCsu。实验分组进行 MTT、凋亡和坏死检测、彗星试验和硫代巴比妥酸(TBARS)、亚硝酸盐、还原型谷胱甘肽(GSH)和过氧化氢酶的反应物质测量。MTT 试验曲线显示 MSCBMsu 和 RPCsu 的平均活力分别为 1.14 ± 0.62 和 1.12 ± 0.54。MSCBMsu 和 RPCsu 的凋亡百分比分别呈现不规则下降和坏死百分比呈现不规则上升的趋势。DNA 损伤指数显示 P5 和 P9 传代的 MSCBMsu 强度较高(<0.05)。在 TBARS 评估中,RPCsu 和 MSCBMsu 的谱系之间存在差异,最后表现出最显著的变化(<0.05)。在亚硝酸盐值方面,我们只在 P1 和 P2 传代的谱系中发现了差异,其中 MSCBMsu 谱系显示出最高的平均值(<0.05)。抗氧化系统活性的测量显示出高标准,仅在 P3 时 RPCsu 谱系中的 GSH 值存在差异(<0.05)。本研究表明,长期维持细胞培养会导致氧化应激的调节降低,并且在体外扩增过程中,RPCsu 比 MSCBMsu 具有更高的遗传稳定性和更低的氧化应激。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/23bf/7368932/d6cdc2775df2/OMCL2020-6470574.001.jpg

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