3P - Single-Cell Isolation of Circulating Tumor Cells by Microfluidic Technology

Date 04 May 2017
Event IMPAKT 2017
Session Welcome reception and Poster Walk
Topics Breast Cancer
Imaging, Diagnosis and Staging
Pathology/Molecular Biology
Presenter Jinho Kim
Authors J. Kim1, H. Cho1, C.W. Jeon2, K. Han1
  • 1Nano Science And Engineering, 197 Inje-ro Gimhae Gyeongsangnam-do Inje University, 50834 - Gimhae/KR
  • 2Department Of Surgery, Gospel Hospital, 262 Gamcheon-ro, Seo-gu, Busan, 49267 - Busan/KR

Abstract

Body

This paper introduces a single-cell isolation microfluidic device for isolating circulating tumor cells from peripheral blood. CTCs from blood are first separated by lateral magnetophoresis. Then, the separated CTCs are electrically identified by single-cell impedance cytometry and isolated as a single-cell by microdispenser.

By recent developments of next-generation sequencing (NGS) and single-cell whole genome amplification (WGA), single-cell genomics is regarded as a core technology for personalized cancer treatment and prophylactic prognosis. In addition, because CTCs are extracted by simple blood draw without pain and are bound to be released from metastasis as well as primary tumor tissues, genomics with single-CTCs is a precious window for understanding genomic diversity of cancer.

For sample preparation, a blood sample is first treated by immunomagnetic nanobeads, specifically binding to MCF-7 breast cancer cells. During sample injection into the SIM-Chip, CTCs in the blood sample are separated by the lateral magnetophoresis and flow into the impedance cytometry. Once the impedance cytometry electrically identifies CTC, the microdispenser propels the shooting buffer in a moment. As a result, a 4.5 µl droplet, containing a single-CTC, is spouted into a collection well through a needle. The percent of droplet containing a single-CTC is 82.4% and the recovery rate was 99.78%, which confirms that the SIM-Chip does not loss CTCs during the isolation of single-CTCs. The viability of isolated single CTCs is 88.53%, compared with 88.98% viability of initial cancer cell lines before the single-cell isolation. That is, the result appeals that the shooting for single-cell isolation does not affect the viability of CTCs. To verify the availability of subsequent genomic assays using the isolated single-CTCs, RT-PCR for detecting KRT19 gene was performed.

In summary, along with recent developed single-cell genomic assay technologies, the presented single-CTC isolation technology will open a new window for understanding genomic diversity of individual patients with cancer, thereby allowing to follow the evolution of cancers and individually tailored cancer treatment.

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