Cancer marker-free enrichment and direct mutation detection in rare cancer cells by combining multi-property isolation and microfluidic concentration

Kim, Soo Hyeon; Ito, Hiroshi; Kozuka, Masahiro; Takagi, Hidenori; Hirai, Mitsuharu; Fujii, Teruo

doi:10.1039/c8lc00772a

Cited by 21 publications

(28 citation statements)

References 29 publications

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“…Due to the complex physics such as strong particle-particle interaction, most of the current microfluidic approaches (e.g., inertial devices) are not applicable for separation in the concentrated sample. Filtration based on porous membranes, including classical membrane filtration 220 and crossflow filtration, 159 is one of the few microsystems that can handle the concentrated sample like whole blood despite their poor recovery rate (<50%) and low throughput. As a result, separation of particles/cells from a highly concentrated biosample remains challenging in the field of microfluidic separation science.…”

Section: Sorting By Shear Induced Diffusionmentioning

confidence: 99%

Label-free microfluidic sorting of microparticles

et al. 2019

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Massive growth of the microfluidics field has triggered numerous advances in focusing, separating, ordering, concentrating, and mixing of microparticles. Microfluidic systems capable of performing these functions are rapidly finding applications in industrial, environmental, and biomedical fields. Passive and label-free methods are one of the major categories of such systems that have received enormous attention owing to device operational simplicity and low costs. With new platforms continuously being proposed, our aim here is to provide an updated overview of the state of the art for passive label-free microparticle separation, with emphasis on performance and operational conditions. In addition to the now common separation approaches using Newtonian flows, such as deterministic lateral displacement, pinched flow fractionation, cross-flow filtration, hydrodynamic filtration, and inertial microfluidics, we also discuss separation approaches using non-Newtonian, viscoelastic flow. We then highlight the newly emerging approach based on shear-induced diffusion, which enables direct processing of complex samples such as untreated whole blood. Finally, we hope that an improved understanding of label-free passive sorting approaches can lead to sophisticated and useful platforms toward automation in industrial, environmental, and biomedical fields.

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Section: Sorting By Shear Induced Diffusionmentioning

confidence: 99%

Label-free microfluidic sorting of microparticles

et al. 2019

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“…In this study, the slit-filter isolation unit of CTC-FIND was used. A filtration module of the slit-filter isolation unit was constructed by sandwiching the slit filter between the sample reservoir and the outlet connected to a tube pump [20].…”

Section: Ctcs Collected By the Slit Filter-based Isolation Of The Ctcmentioning

confidence: 99%

“…In our previous study, we proposed a method of CTC enrichment by size-based filtration and immunomagnetic negative selection followed by dielectrophoretic concentration (CTC-FIND) [20]. The high purity and high recovery rate of this system were verified using a spike-in experiment.…”

Section: Introductionmentioning

confidence: 99%

Analysis of the Circulating Tumor Cell Capture Ability of a Slit Filter-Based Method in Comparison to a Selection-Free Method in Multiple Cancer Types

Takagi

Dong

Kuczler

et al. 2020

IJMS

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Circulating tumor cells (CTCs) are a promising biomarker for cancer liquid biopsy. To evaluate the CTC capture bias and detection capability of the slit filter-based CTC isolation platform (CTC-FIND), we prospectively compared it head to head to a selection-free platform (AccuCyte®-CyteFinder® system). We used the two methods to determine the CTC counts, CTC positive rates, CTC size distributions, and CTC phenotypes in 36 patients with metastatic cancer. Between the two methods, the median CTC counts were not significantly different and the total counts were correlated (r = 0.63, p < 0.0001). The CTC positive rate by CTC-FIND was significantly higher than that by AccuCyte®-CyteFinder® system (91.7% vs. 66.7%, p < 0.05). The median diameter of CTCs collected by CTC-FIND was significantly larger (13.0 μm, range 5.2–52.0 vs. 10.4 μm, range 5.2–44.2, p < 0.0001). The distributions of CTC phenotypes (CK+EpCAM+, CK+EpCAM− or CK−EpCAM+) detected by both methods were similar. These results suggested that CTC-FIND can detect more CTC-positive cases but with a bias toward large size of CTCs.

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“…We recently developed CTC-FIND which is a technique combining filter separation with immunomagnetic depletion of CD45/50-expressing cells to isolate ultra-pure CTCs without relying on any traditional CTC markers, such as epithelial and mesenchymal cell antigens, and validated by blood samples collected from healthy volunteers [ 17 ]. By using CTC-FIND, we previously demonstrated that this method enabled us to not only collect typical epithelial cancer cells but also non-epithelial cancer cells with a residual of only about a hundred leucocytes from 8 mL whole blood.…”

Section: Introductionmentioning

confidence: 99%

Clinical Significance of Circulating Tumor Cell Induced Epithelial-Mesenchymal Transition in Patients with Metastatic Colorectal Cancer by Single-Cell RNA-Sequencing

Kozuka

Battaglin

Jayachandran

et al. 2021

Cancers

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Background: Circulating tumor cells (CTCs) are a prognostic marker in patients with metastatic colorectal cancer (mCRC). However, little is known about the characterization of CTCs in mCRC at the single-cell level using RNA sequencing. The purpose of this study was to validate the capability to detect and isolate single CTCs for single-cell RNA sequencing (scRNA-seq) and to identify clinical significance at a single CTC level. Methods: Single CTCs from 27 mCRC patients were collected by CTC-FIND, which is comprised of filter separation and immunomagnetic depletion to collect ultra-pure CTC samples. To address tumor heterogeneity, CTCs were collected without relying on any traditional CTC markers, such as epithelial and mesenchymal cell antigens, and were undertaken by scRNA-seq using SMART-Seq v4. Results: We identified 59 single CTCs which were classified into four groups by epithelial, epithelial-mesenchymal transition (EMT) and stem cell-related gene expression. Patients receiving second or later-line treatment who had EMT gene expressing CTCs had a significantly shorter PFS and OS. Conclusions: Exploiting CTC-FIND with SMART-Seq v4 showed that scRNA-seq of CTCs may shed new insight into tumor heterogeneity of mCRC and that the presence of CTCs expressing EMT-related genes at the single-cell level could have prognostic value in mCRC patients.

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Cancer marker-free enrichment and direct mutation detection in rare cancer cells by combining multi-property isolation and microfluidic concentration

Abstract: We present a novel cancer marker-free CTC enrichment method by size-based filtration and immunomagnetic negative selection followed by dielectrophoretic concentration for direct detection of genetic mutations in rare cancer cells suspended in whole blood.

Cited by 21 publications

References 29 publications

Label-free microfluidic sorting of microparticles

Label-free microfluidic sorting of microparticles

Analysis of the Circulating Tumor Cell Capture Ability of a Slit Filter-Based Method in Comparison to a Selection-Free Method in Multiple Cancer Types

Clinical Significance of Circulating Tumor Cell Induced Epithelial-Mesenchymal Transition in Patients with Metastatic Colorectal Cancer by Single-Cell RNA-Sequencing

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