Abstract：
In vitro biofilm models have allowed researchers to investigate the role biofilms play in the pathogenesis, virulence, and antimicrobial drug susceptibility of a wide range of bacterial pathogens. Rotary cell culture systems create three-dimensional cellular structures, primarily applied to eukaryotic organoids, that better capture characteristics of the cells in vivo. Here, we describe how to apply a low-shear, detergent-free rotary cell culture system to generate biofilms of Mycobacterium bovis BCG. The three-dimensional biofilm model forms mycobacterial cell aggregates in suspension as surface-detached biomass, without severe nutrient starvation or environmental stress, that can be harvested for downstream experiments. Mycobacterium bovis BCG derived from cell clusters display antimicrobial drug tolerance, presence of an extracellular matrix, and evidence of cell wall remodeling, all features of biofilm-associated bacteria that may be relevant to the treatment of tuberculosis.
摘要：
体外生物膜模型使研究人员能够研究生物膜在发病机理中的作用，毒力,和抗菌药物敏感性广泛的细菌病原体。旋转细胞培养系统创建三维细胞结构，主要应用于真核细胞，更好地捕获体内细胞的特性。这里,我们描述了如何应用低剪切，无去污剂的旋转细胞培养系统，以产生牛分枝杆菌BCG的生物膜。三维生物膜模型在悬浮液中形成分枝杆菌细胞聚集体作为表面分离的生物质，没有严重的营养饥饿或环境压力，可以收获用于下游实验。来自细胞簇的牛分枝杆菌BCG显示抗菌药物耐受性，细胞外基质的存在，以及细胞壁重塑的证据，可能与结核病治疗相关的生物膜相关细菌的所有特征。
