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Mycoplasma contamination of leukemic cell lines alters protein expression determined by reverse phase protein arrays

机译:白血病细胞系的支原体污染改变了由反相蛋白质阵列测定的蛋白质表达

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摘要

Mycoplasma contamination is a major problem in cell culturing, potentially altering the results of cell line-based experiments in largely uncharacterized ways. To define the consequences of mycoplasma infection at the level of protein expression we utilized the reverse phase protein array technology to analyze the expression of 235 proteins in mycoplasma infected, uninfected post treatment, and never-infected leukemic cell lines. Overall, protein profiles of cultured cells remained relatively stable after mycoplasma infection. However, paired comparisons for individual proteins identified that 18.7% of the proteins significantly changed between the infected and the never-infected cell line samples, and that 14.0% of the proteins significantly altered between the infected and the post treatment samples. Six percent of the proteins were affected in the post treatment samples compared to the never-infected samples, and 7.2% compared to treated cells that had never had mycoplasma infection before. Proteins that were significantly altered in the infected cells were enriched for apoptotic signaling processes and auto-phosphorylation, suggesting an increased cellular stress and a decreased growth rate. In conclusion, this study shows that mycoplasma infection of leukemic cell lines alters the proteins expression levels, potentially confounding experimental results. This reinforces the need for regular testing of mycoplasma. Electronic supplementary material The online version of this article (10.1007/s10616-018-0244-2) contains supplementary material, which is available to authorized users.
机译:支原体污染是细胞培养中的一个主要问题,可能会以许多未知的方式改变基于细胞系的实验结果。为了确定支原体感染在蛋白质表达水平上的后果,我们利用反相蛋白质阵列技术分析了感染,未感染的后处理和从未感染的白血病细胞系中235种蛋白质的表达。总体而言,支原体感染后培养细胞的蛋白质谱保持相对稳定。然而,对单个蛋白质的配对比较发现,感染和从未感染的细胞系样本之间有18.7%的蛋白质发生了显着变化,感染和未处理的样本之间有14.0%的蛋白质发生了显着变化。与从未感染的样本相比,在处理后样本中有6%的蛋白质受到了影响,与之前从未感染过支原体的处理过的细胞相比,有7.2%的蛋白质受到了影响。感染细胞中发生显着变化的蛋白质富含凋亡信号转导过程和自身磷酸化,提示细胞压力增加和生长速率降低。总之,这项研究表明,白血病细胞株的支原体感染会改变蛋白质的表达水平,可能会混淆实验结果。这增加了定期检测支原体的需要。电子补充材料本文的在线版本(10.1007 / s10616-018-0244-2)包含补充材料,授权用户可以使用。

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