Institute for Breast Health Medicine, State Key Laboratory of Biotherapy, West China Hospital, Sichuan University, Chengdu, Sichuan 610041, China.
Department of Pancreatic Surgery, West China Hospital, Sichuan University, Chengdu, Sichuan 610041, China.
STAR Protoc. 2023 Sep 15;4(3):102539. doi: 10.1016/j.xpro.2023.102539. Epub 2023 Sep 5.
Circulating tumor cells (CTCs) are regarded as the "seeds" of tumor metastasis. Identifying immune checkpoints on CTCs is essential for establishing efficient immunotherapies to prevent tumor metastasis. Here, we present a protocol for isolating CTCs and obtaining single-cell suspensions from pancreatic ductal adenocarcinoma liver metastatic patients. We describe steps for biospecimen acquisition, CTC isolation, and tissue dissociation. We then detail procedures for performing single-cell RNA-seq, annotating cell types, and identifying immune checkpoints on CTCs. For complete details on the use and execution of this protocol, please refer to Liu et al. (2023)..
循环肿瘤细胞 (CTCs) 被认为是肿瘤转移的“种子”。鉴定 CTC 上的免疫检查点对于建立有效的免疫疗法以防止肿瘤转移至关重要。在这里，我们提供了一种从胰腺导管腺癌肝转移患者中分离 CTC 并获得单细胞悬液的方案。我们描述了生物样本采集、CTC 分离和组织解离的步骤。然后，我们详细介绍了进行单细胞 RNA-seq、注释细胞类型和鉴定 CTC 上免疫检查点的程序。有关此方案的使用和执行的完整详细信息，请参阅 Liu 等人。(2023)。
