循环肿瘤细胞
液体活检
生物
全血
数字聚合酶链反应
癌症研究
癌症
计算生物学
病理
医学
免疫学
基因
聚合酶链反应
转移
遗传学
作者
Risa Burr,Jon F.,Brian Chirn,Avanish Mishra,Daniel A. Haber,Mehmet Toner,Shyamala Maheswaran
出处
期刊:Methods in molecular biology
日期:2022-01-01
卷期号:: 309-321
被引量:2
标识
DOI:10.1007/978-1-0716-2193-6_18
摘要
The ability to isolate and analyze rare circulating tumor cells (CTCs) holds the potential to increase our understanding of cancer evolution and allows monitoring of disease and therapeutic responses through a relatively non-invasive blood-based biopsy. While many methods have been described to isolate CTCs from the blood, the vast majority rely on size-based sorting or positive selection of CTCs based on surface markers, which introduces bias into the downstream product by making assumptions about these heterogenous cells. Here we describe a negative-selection protocol for enrichment of CTCs through removal of blood components including red blood cells, platelets, and white blood cells. This procedure results in a product that is amenable to downstream single-cell analytics including RNA-Seq, ATAC-Seq and DNA methylation, droplet digital PCR (ddPCR) for tumor specific transcripts, staining and extensive image analysis, and ex vivo culture of patient-derived CTCs.
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