Tracking and characterization of partial and full epithelial-mesenchymal transition cells in a mouse model of metastatic breast cancer

• Published in: STAR protocols Vol. 3; no. 2; p. 101438
• Main Authors: Lüönd, Fabiana, Santacroce, Natascha, Beisel, Christian, Guérard, Laurent, Bürglin, Thomas R., Christofori, Gerhard, Sugiyama, Nami
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 17.06.2022; Elsevier
• Subjects: Cancer; Cell Biology; Cell Differentiation; Flow Cytometry/Mass Cytometry; Microscopy; Molecular Biology; Protocol; RNAseq; Single Cell; Microscopy; Molecular Biology; Flow Cytometry/Mass Cytometry; Single Cell; Cell Differentiation; RNAseq; Cell Biology; Cancer
• ISSN: 2666-1667; 2666-1667
• DOI: 10.1016/j.xpro.2022.101438

The various stages of epithelial-mesenchymal transition (EMT) generate phenotypically heterogeneous populations of cells. Here, we detail a dual recombinase lineage tracing system using a transgenic mouse model of metastatic breast cancer to trace and characterize breast cancer cells at different EMT stages. We describe analytical steps to label cancer cells at an early partial or a late full EMT state, followed by tracking their behavior in tumor slice cultures. We then characterize their transcriptome by five-cell RNA sequencing. For complete details on the use and execution of this protocol, please refer to Luond et al. (2021). [Display omitted] •A dual recombinase lineage tracing system for cancer cells undergoing EMT•Enables live imaging of cancer cells undergoing early or late EMT•Supports flow-cytometry-mediated isolation of cancer cells in various stages of EMT•Enables single-cell transcriptomic studies of cancer cells undergoing EMT Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. The various stages of epithelial-mesenchymal transition (EMT) generate phenotypically heterogeneous populations of cells. Here, we detail a dual recombinase lineage tracing system using a transgenic mouse model of metastatic breast cancer to trace and characterize breast cancer cells at different EMT stages. We describe analytical steps to label cancer cells at an early partial or a late full EMT state, followed by tracking their behavior in tumor slice cultures. We then characterize their transcriptome by five-cell RNA sequencing.