Amyloid-beta peptide degradation in cell cultures by mycoplasma contaminants.
Amyloid-beta peptide degradation in cell cultures by mycoplasma contaminants.
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DOI：
10.1186/1756-0500-1-38
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发表时间：
2008-06-30
影响因子：
1.8
通讯作者：
Marambaud P
中科院分区：
文献类型：
--
作者：
Zhao H;Dreses-Werringloer U;Davies P;Marambaud P
细胞培养已成为阿尔茨海默病研究中不可或缺的研究β淀粉样蛋白（Aβ）代谢的工具。据估计，目前使用的高达35%的细胞培养物被各种支原体感染。与常见的细菌和真菌感染相比，支原体对细胞培养物的污染在可检测性和预防方面是一个具有挑战性的问题。支原体是最小和最简单的自我复制细菌，感染宿主细胞的后果是可变的，从没有明显的影响到诱导细胞凋亡。在这里，我们提出的证据表明，来自细胞培养污染的支原体能够有效和快速地降解细胞外Aβ。结果，我们在稳定转染淀粉样β前体蛋白（APP）的支原体阳性细胞的条件培养基中未观察到Aβ蓄积。重要的是，根除支原体污染物-确定为M。hyorhinis -通过用基于喹诺酮的抗生素处理，在APP转染的细胞中恢复了细胞外Aβ积累。这些数据表明，支原体降解Aβ，因此在比较不同细胞系中的细胞外Aβ水平时，可能是变异性的重要来源。基于这些结果，我们建议在培养细胞中测量细胞外Aβ水平之前评估支原体污染。
Cell cultures have become an indispensable tool in Alzheimer's disease research for studying amyloid-β (Aβ) metabolism. It is estimated that up to 35% of cell cultures in current use are infected with various mycoplasma species. In contrast with common bacterial and fungal infections, contaminations of cell cultures with mycoplasmas represent a challenging issue in terms of detectability and prevention. Mycoplasmas are the smallest and simplest self-replicating bacteria and the consequences of an infection for the host cells are variable, ranging from no apparent effect to induction of apoptosis. Here we present evidence that mycoplasmas from a cell culture contamination are able to efficiently and rapidly degrade extracellular Aβ. As a result, we observed no accumulation of Aβ in the conditioned medium of mycoplasma-positive cells stably transfected with the amyloid-β precursor protein (APP). Importantly, eradication of the mycoplasma contaminant – identified as M. hyorhinis – by treatments with a quinolone-based antibiotic, restored extracellular Aβ accumulation in the APP-transfected cells. These data show that mycoplasmas degrade Aβ and thus may represent a significant source of variability when comparing extracellular Aβ levels in different cell lines. On the basis of these results, we recommend assessment of mycoplasma contaminations prior to extracellular Aβ level measurements in cultured cells.