Department of Biomedicine, University of Basel, 4058 Basel, Switzerland.
Department of Biosystems Science and Engineering, ETH Zürich, 4058 Basel, Switzerland.
STAR Protoc. 2022 Jun 10;3(2):101438. doi: 10.1016/j.xpro.2022.101438. eCollection 2022 Jun 17.
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).
上皮-间充质转化 (EMT) 的各个阶段产生表型异质的细胞群体。在这里，我们详细描述了一种使用转移性乳腺癌转基因小鼠模型的双重组酶谱系追踪系统，以追踪和表征不同 EMT 阶段的乳腺癌细胞。我们描述了标记处于早期部分 EMT 或晚期完全 EMT 状态的癌细胞的分析步骤，然后追踪它们在肿瘤切片培养物中的行为。然后，我们通过五细胞 RNA 测序对其转录组进行特征分析。有关该协议的使用和执行的完整详细信息，请参阅 Luond 等人。（2021 年）。
