1. PIC&RUN: An integrated assay for the detection and retrieval of single viable circulating tumor cells
- Author
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Oihana Iriondo, Mohamed Kamal, Yonatan Amzaleg, Shahin Saremi, Irene Kang, Varsha Tulpule, Julie E. Lang, Remi Klotz, Min Yu, and Yvonne Chairez
- Subjects
Adult ,0301 basic medicine ,Computer science ,lcsh:Medicine ,Breast Neoplasms ,Cell Count ,Immune markers ,Cell Separation ,Computational biology ,Cellular level ,Article ,Metastasis ,03 medical and health sciences ,Negative selection ,Breast cancer ,0302 clinical medicine ,Circulating tumor cell ,Biological property ,Biomarkers, Tumor ,Tumor Cells, Cultured ,Humans ,Liquid biopsy ,Author Correction ,lcsh:Science ,Aged ,Multidisciplinary ,Gene Expression Profiling ,Positive selection ,lcsh:R ,Disease progression ,Middle Aged ,Neoplastic Cells, Circulating ,Gene Expression Regulation, Neoplastic ,030104 developmental biology ,Case-Control Studies ,030220 oncology & carcinogenesis ,Female ,lcsh:Q ,Single-Cell Analysis - Abstract
Circulating tumor cells (CTCs) shed from solid tumors can serve as a minimally invasive liquid biopsy for monitoring disease progression. Because CTCs are rare and heterogeneous, their biological properties need to be investigated at the single cell level, which requires efficient ways to isolate and analyze live single CTCs. Current methods for CTC isolation and identification are either performed on fixed and stained cells or need multiple procedures to isolate pure live CTCs. Here, we used the AccuCyte-RareCyte system to develop a Protocol for Integrated Capture and Retrieval of Ultra-pure single live CTCs using Negative and positive selection (PIC&RUN). The positive selection module of PIC&RUN identifies CTCs based on detection of cancer surface markers and exclusion of immune markers. Combined with a two-step cell picking protocol to retrieve ultrapure single CTCs, the positive selection module is compatible for downstream single cell transcriptomic analysis. The negative selection module of PIC&RUN identifies CTCs based on a live cell dye and the absence of immune markers, allowing retrieval of viable CTCs that are suitable for ex vivo culture. This new assay combines the CTC capture and retrieval in one integrated platform, providing a valuable tool for downstream live CTC analyses.
- Published
- 2019