Isolation and genome sequencing of individual circulating tumor cells using hydrogel encapsulation and laser capture microdissection

Lab on a Chip
Emily S ParkHongshen Ma

Abstract

Circulating tumor cells (CTCs) are malignant cells released into the bloodstream with the potential to form metastases in secondary sites. These cells, acquired non-invasively, represent a sample of highly relevant tumor tissue that is an alternative to difficult and low-yield tumor biopsies. In recent years, there has been growing interest in genomic profiling of CTCs to enable longitudinal monitoring of the tumor's adaptive response to therapy. However, due to their extreme rarity, genotyping CTCs has proved challenging. Relevant mutations can be masked by leukocyte contamination in isolates. Heterogeneity between subpopulations of tumor cells poses an additional obstacle. Recent advances in single-cell sequencing can overcome these limitations but isolation of single CTCs is prone to cell loss and is prohibitively difficult and time consuming. To address these limitations, we developed a single cell sample preparation and genome sequencing pipeline that combines biophysical enrichment and single cell isolation using laser capture microdissection (LCM). A key component of this process is the encapsulation of enriched CTC sample in a hydrogel matrix, which enhances the efficiency of single-cell isolation by LCM, and is compati...Continue Reading

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Citations

Oct 9, 2019·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Mingxin XuLichuan Zhang
Aug 29, 2019·Nature Reviews. Cancer·Laura Keller, Klaus Pantel
Jan 25, 2020·Journal of Clinical Medicine·Nak Jun LeeKi Wan Bong
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Jan 16, 2021·Molecular Oncology·Mahmoud Labib, Shana O Kelley
Sep 26, 2021·Journal of Basic and Clinical Physiology and Pharmacology·Felice CrocettoDaniela Terracciano

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Methods Mentioned

BETA
biopsies
flow cytometry
genotyping
fluorescence microscopy
electrophoresis
PCR
Illumina sequencing
single cell sequencing

Software Mentioned

SAMtools
AdhesiveCap
Bowtie
Chromas

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