Division of Pulmonary Allergy Critical Care and Sleep Medicine, Davis Heart and Lung Research Institute, The Ohio State University, Columbus, Ohio, United States of America.
PLoS One. 2010 Mar 25;5(3):e9900. doi: 10.1371/journal.pone.0009900.
Mycoplasma contamination of cultured cell lines is a serious problem in research, altering cellular response to different stimuli thus compromising experimental results. We found that chronic mycoplasma contamination of THP-1 cells suppresses responses of THP-1 cells to TLR stimuli. For example, E. coli LPS induced IL-1 beta was suppressed by 6 fold and IL-8 by 10 fold in mycoplasma positive THP-1 cells. Responses to live F. novicida challenge were suppressed by 50-fold and 40-fold respectively for IL-1beta and IL-8. Basal TLR4 expression level in THP-1 cells was decreased by mycoplasma by 2.4-fold (p = 0.0003). Importantly, cell responses to pathogen associated molecular patterns are completely restored by mycoplasma clearance with Plasmocin. Thus, routine screening of cell lines for mycoplasma is important for the maintenance of reliable experimental data and contaminated cell lines can be restored to their baseline function with antibiotic clearance of mycoplasma.
支原体污染是细胞培养中的一个严重问题，它会改变细胞对不同刺激的反应，从而影响实验结果。我们发现，THP-1 细胞的慢性支原体污染会抑制 THP-1 细胞对 TLR 刺激的反应。例如，在支原体阳性的 THP-1 细胞中，大肠杆菌 LPS 诱导的 IL-1β被抑制了 6 倍，IL-8 被抑制了 10 倍。对活的 F. novicida 挑战的反应分别被抑制了 50 倍和 40 倍，用于 IL-1β和 IL-8。THP-1 细胞中 TLR4 的基础表达水平被支原体降低了 2.4 倍（p = 0.0003）。重要的是，用 Plasmocin 清除支原体可以完全恢复细胞对病原体相关分子模式的反应。因此，常规筛选细胞系中的支原体对于维护可靠的实验数据非常重要，并且可以通过清除支原体来恢复被污染的细胞系的基线功能。
