Single-Cell Isolation of Circulating Tumor Cells from Whole Blood by Lateral Magnetophoretic Microseparation and Microfluidic Dispensing

Kim, Jin-Ho; Cho, Hyungseok; Han, Song‐I; Han, Ki-Ho

doi:10.1021/acs.analchem.6b00570

Cited by 66 publications

(37 citation statements)

References 37 publications

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“…Kim et al . used an air pressure driven dispensing controller to shoot out single circulating tumor cells through an exhaust needle29. However, the needle was arranged vertically to the fluid channel and in parallel to the objective, thus the single-cell exporting procedure could not be visualized, which might lead to imprecise collection.…”

Section: Resultsmentioning

confidence: 99%

Development of a facile droplet-based single-cell isolation platform for cultivation and genomic analysis in microorganisms

Zhang

Wang

Zhou

et al. 2017

Sci Rep

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Wider application of single-cell analysis has been limited by the lack of an easy-to-use and low-cost strategy for single-cell isolation that can be directly coupled to single-cell sequencing and single-cell cultivation, especially for small-size microbes. Herein, a facile droplet microfluidic platform was developed to dispense individual microbial cells into conventional standard containers for downstream analysis. Functional parts for cell encapsulation, droplet inspection and sorting, as well as a chip-to-tube capillary interface were integrated on one single chip with simple architecture, and control of the droplet sorting was achieved by a low-cost solenoid microvalve. Using microalgal and yeast cells as models, single-cell isolation success rate of over 90% and single-cell cultivation success rate of 80% were demonstrated. We further showed that the individual cells isolated can be used in high-quality DNA and RNA analyses at both gene-specific and whole-genome levels (i.e. real-time quantitative PCR and genome sequencing). The simplicity and reliability of the method should improve accessibility of single-cell analysis and facilitate its wider application in microbiology researches.

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Section: Resultsmentioning

confidence: 99%

Development of a facile droplet-based single-cell isolation platform for cultivation and genomic analysis in microorganisms

Zhang

Wang

Zhou

et al. 2017

Sci Rep

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“…The combined enrichment technologies are termed as dual modality enrichment, such as density gradient sedimentation plus size exclusion filtration, 49 and microfluidic chips plus magnetic beads. 50 …”

Section: Ctc Detection Technologies With Enrichmentmentioning

confidence: 99%

“…50 The SIM-Chip comprises a lateral magnetophoretic microseparator and a microdispenser as a two-step cascade platform. First, CTCs were enriched from whole blood by the lateral magnetophoretic microseparator based on immunomagnetic nanobeads.…”

Section: Ctc Detection Technologies With Enrichmentmentioning

confidence: 99%

Current detection technologies for circulating tumor cells

2017

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Circulating tumor cells (CTCs) are cancer cells that circulate in the blood stream after being naturally shed from original or metastatic tumors, and can lead to a new fatal metastasis. CTCs have become a hotspot research field during the last decade. Detection of CTCs, as a liquid biopsy of tumors, can be used for early diagnosis of cancers, earlier evaluation of cancer recurrence and chemotherapeutic efficacy, and choice of individual sensitive anti-cancer drugs. Therefore, CTC detection is a crucial tool to fight against cancer. Herein, we classify the currently reported CTC detection technologies, introduce some representative samples for each technology, conclude the advantages and limitations, and give a future prospective including challenges and opportunities of CTC detection.

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“…Currently, there are few techniques available for single cell isolation, including micromanipulation, laser micro-dissection and high throughput fluorescence-activated cell sorting (FACS). These approaches have several disadvantages including inadequate detection sensitivity for CTC population, a required high number of cells as a starting population and are manual and laborious methods [28,29,30]. …”

Section: Introductionmentioning

confidence: 99%

Detection of Activating Estrogen Receptor Gene (ESR1) Mutations in Single Circulating Tumor Cells

Paolillo

Rossi

et al. 2017

Clinical Cancer Research

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Purpose Early detection is essential for treatment plans before onset of metastatic disease. Our purpose was to demonstrate feasibility to detect and monitor estrogen receptor 1 (ESR1) gene mutations at the single circulating tumor cell (CTC) level in metastatic breast cancer (MBC). Experimental Design We used a CTC molecular characterization approach to investigate heterogeneity of 14 hot spot mutations in ESR1 and their correlation with endocrine resistance. Combining the CellSearch® and DEPArray™ technologies allowed recovery of 71 single CTCs and 12 WBC from 3 ER-positive MBC patients. 40 CTCs and 12 WBC were subjected to whole genome amplification by MALBAC and Sanger sequencing. Results Among 3 selected patients, 2 had an ESR1 mutation (Y537). One showed two different ESR1 variants in a single CTC and another showed loss of heterozygosity. All mutations were detected in matched cell-free DNA (cfDNA). Furthermore, one had 2 serial blood samples analyzed and showed changes in both cfDNA and CTCs with emergence of mutations in ESR1 (Y537S and T570I), which has not been reported previously. Conclusions CTCs are easily accessible biomarkers to monitor and better personalize management of patients with previously demonstrated ER-MBC who are progressing on endocrine therapy. We showed that single CTC analysis can yield important information on clonal heterogeneity, and can be a source of discovery of novel and potential driver mutations. Finally, we also validate a workflow for liquid biopsy that will facilitate early detection of ESR1 mutations, the emergence of endocrine resistance and the choice of further target therapy.

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Single-Cell Isolation of Circulating Tumor Cells from Whole Blood by Lateral Magnetophoretic Microseparation and Microfluidic Dispensing

Cited by 66 publications

References 37 publications

Development of a facile droplet-based single-cell isolation platform for cultivation and genomic analysis in microorganisms

Development of a facile droplet-based single-cell isolation platform for cultivation and genomic analysis in microorganisms

Current detection technologies for circulating tumor cells

Detection of Activating Estrogen Receptor Gene (ESR1) Mutations in Single Circulating Tumor Cells

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