Summary Links between bacterial infections and cancer are actively investigated. Cost-effective assays to quantify bacterial oncogenic potential can shed new light on these links. Here, we present a soft agar colony formation assay to quantify mouse embryonic fibroblast transformation after Salmonella Typhimurium infection. We describe how to infect and seed cells in soft agar for anchorage-independent growth, a hallmark of cell transformation. We further detail automated cell colony enumeration. This protocol is adaptable to other bacteria or host cells. For complete details on the use and execution of this protocol, please refer to Van Elsland et al. 1 Graphical abstract Highlights • Infection of mouse embryonic fibroblasts (MEF) with Salmonella Typhimurium • Seeding MEF post-infection in soft agar for anchorage-independent growth assay • Automated acquisition and counting of MEF colonies formed in soft agar • Cost-effective assessment of cell transformation upon bacterial infection Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Links between bacterial infections and cancer are actively investigated. Cost-effective assays to quantify bacterial oncogenic potential can shed new light on these links. Here, we present a soft agar colony formation assay to quantify mouse embryonic fibroblast transformation after Salmonella Typhimurium infection. We describe how to infect and seed cells in soft agar for anchorage-independent growth, a hallmark of cell transformation. We further detail automated cell colony enumeration. This protocol is adaptable to other bacteria or host cells.
【저자키워드】 Microbiology, Cancer, Cell culture,