体外生物膜模型使研究人员能够研究生物膜在多种细菌病原体的发病机制、毒力和抗菌药物敏感性中所起的作用。旋转细胞培养系统创建三维细胞结构，主要应用于真核类器官，可以更好地捕获体内细胞的特征。在这里，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来生成牛分枝杆菌 BCG 生物膜。三维生物膜模型形成悬浮的分枝杆菌细胞聚集体作为表面分离的生物质，没有严重的营养缺乏或环境压力，可以收获用于下游实验。源自细胞簇的牛分枝杆菌 BCG 显示出抗菌药物耐受性、细胞外基质的存在以及细胞壁重塑的证据，这些生物膜相关细菌的所有特征可能与结核病的治疗相关。© 2024。作者，获得 Springer Science Business Media, LLC（Springer Nature 旗下公司）的独家许可。

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.© 2024. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.