Protocol for identifying immune checkpoint on circulating tumor cells of human pancreatic ductal adenocarcinoma by single-cell RNA sequencing

被引：3

作者：

Liu, Xiaowei ^{[1
]}

Song, Jinen ^{[1
]}

Liu, Xinyu ^{[1
]}

Zhang, Hao ^{[4
]}

Wang, Xueyan ^{[1
]}

Li, Yuanxi ^{[1
]}

Yang, Zhankun ^{[5
]}

Jing, Jing ^{[1
]}

Ma, Xuelei ^{[2
,3
]}

Shi, Hubing ^{[1
]}

机构：

[1] Sichuan Univ, West China Hosp, Inst Breast Hlth Med, State Key Lab Biotherapy, Chengdu 610041, Sichuan, Peoples R China

[2] Sichuan Univ, West China Hosp, Dept Biotherapy, Chengdu 610041, Sichuan, Peoples R China

[3] Sichuan Univ, State Key Lab Biotherapy, Chengdu 610041, Sichuan, Peoples R China

[4] Sichuan Univ, West China Hosp, Dept Pancreat Surg, Chengdu 610041, Sichuan, Peoples R China

[5] Shijiazhuang Univ, Coll Chem Engn, Shijiazhuang 050035, Hebei, Peoples R China

来源：

STAR PROTOCOLS | 2023年 / 4卷 / 03期

基金：

中国国家自然科学基金;

关键词：

Bioinformatics; Cancer; Cell Biology; Cell Isolation; Flow Cytometry/Mass Cytometry; Genomics; Health Sciences; High Throughput Screening; Immunology; RNA-seq; Sequence Analysis; Sequencing; Single Cell;

D O I：

10.1016/j.xpro.2023.102539

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

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).1

引用

页数：26

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