Zhao Haitian, Dreses-Werringloer Ute, Davies Peter, Marambaud Philippe
Litwin-Zucker Research Center for the Study of Alzheimer Disease, The Feinstein Institute for Medical Research, North Shore-LIJ, Manhasset, NY, USA.
BMC Res Notes. 2008 Jun 30;1:38. doi: 10.1186/1756-0500-1-38.
Cell cultures have become an indispensable tool in Alzheimer's disease research for studying amyloid-beta (Abeta) 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 Abeta. As a result, we observed no accumulation of Abeta in the conditioned medium of mycoplasma-positive cells stably transfected with the amyloid-beta precursor protein (APP). Importantly, eradication of the mycoplasma contaminant - identified as M. hyorhinis - by treatments with a quinolone-based antibiotic, restored extracellular Abeta accumulation in the APP-transfected cells.
These data show that mycoplasmas degrade Abeta and thus may represent a significant source of variability when comparing extracellular Abeta levels in different cell lines. On the basis of these results, we recommend assessment of mycoplasma contaminations prior to extracellular Abeta level measurements in cultured cells.
细胞培养已成为阿尔茨海默病研究中研究β淀粉样蛋白(Aβ)代谢不可或缺的工具。据估计,目前使用的细胞培养物中高达35%受到各种支原体物种的感染。与常见的细菌和真菌感染不同,支原体污染细胞培养物在可检测性和预防方面是一个具有挑战性的问题。支原体是最小且最简单的自我复制细菌,感染对宿主细胞的影响各不相同,从无明显影响到诱导细胞凋亡。
我们在此提供证据表明,细胞培养物污染中的支原体能够高效、快速地降解细胞外Aβ。因此,我们观察到稳定转染淀粉样前体蛋白(APP)的支原体阳性细胞的条件培养基中没有Aβ积累。重要的是,通过使用喹诺酮类抗生素治疗根除被鉴定为猪鼻支原体的支原体污染物后,APP转染细胞中的细胞外Aβ积累得以恢复。
这些数据表明,支原体可降解Aβ,因此在比较不同细胞系中的细胞外Aβ水平时可能是一个显著的变异性来源。基于这些结果,我们建议在测量培养细胞中的细胞外Aβ水平之前评估支原体污染情况。