1 Laboratory of Microbiology, Wageningen University, Dreijenplein 10, 6703 HB Wageningen, the Netherlands.
2 Department of Veterinary Biosciences, University of Helsinki, P.O. Box 66, 00014 Helsinki, Finland.
Benef Microbes. 2017 Apr 26;8(2):163-169. doi: 10.3920/BM2016.0096. Epub 2017 Jan 24.
The anaerobic gut bacterium Akkermansia muciniphila is a well-characterised member of the mucosal microbiota and has shown to be a gut symbiont in human. A. muciniphila has been negatively associated with obesity and its associated metabolic disorders in various human cohorts while treatment with A. muciniphila cells reversed highfat diet-induced obesity and its associated metabolic disorders in mouse models. Therefore, administration of A. muciniphila has been suggested as a possible new therapeutic treatment for these omnipresent diseases. Here we describe a potentially scalable workflow for the preparation and preservation of high numbers of viable cells of A. muciniphila obtained from 1 l laboratory scale growth under strict anaerobic conditions for therapeutic interventions. This resulted in viable A. muciniphila cells with high yields and very high stability, with up to 97.9±4.5% survival for a time period of 1 year at -80 °C in glycerol-amended medium. Moreover, various quality assessment and control procedures were developed to ensure the use of viable cells of A. muciniphila. Several microscopic, culturing, and molecular approaches were applied to monitor the presence, abundance and recovery of A. muciniphila before, during, and after its administration to high-fat treated mice. We show that viable A. muciniphila cells can be recovered from caecal and colon content (up to 1×10 cells/g), testifying for the efficiency of the described workflow.
厌氧肠道细菌阿克曼氏菌（Akkermansia muciniphila）是黏膜微生物群的一个特征成员，已被证明是人类肠道共生体。在各种人类队列中，A. muciniphila 与肥胖及其相关代谢紊乱呈负相关，而用 A. muciniphila 细胞治疗可逆转高脂肪饮食诱导的肥胖及其在小鼠模型中的相关代谢紊乱。因此，A. muciniphila 的给药已被建议作为这些普遍存在的疾病的一种可能的新治疗方法。在这里，我们描述了一种潜在可扩展的工作流程，用于在严格厌氧条件下从 1 升实验室规模生长中制备和保存大量可存活的 A. muciniphila 细胞，用于治疗干预。这导致了可存活的 A. muciniphila 细胞具有高产率和非常高的稳定性，在甘油改良培养基中 -80°C 下保存长达 1 年，存活率高达 97.9±4.5%。此外，还开发了各种质量评估和控制程序，以确保使用可存活的 A. muciniphila 细胞。应用了几种显微镜、培养和分子方法来监测 A. muciniphila 在给予高脂肪治疗的小鼠之前、期间和之后的存在、丰度和恢复情况。我们表明，可存活的 A. muciniphila 细胞可以从盲肠和结肠内容物（高达 1×10 个细胞/g）中回收，证明了所描述的工作流程的效率。
