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Micromachines
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Microfluidics for Circulating Biomarkers
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Microfluidics for Circulating Biomarkers

A special issue of Micromachines (ISSN 2072-666X). This special issue belongs to the section "B：Biology and Biomedicine".

Deadline for manuscript submissions: closed (15 October 2018) | Viewed by 51145

Special Issue Editor

Prof. Dr. Hyo-il Jung

E-Mail Website
Guest Editor
Laboratory of Biochip Technology, School of Mechanical Engineering, Yonsei University, 50 Yonsei-no Seodaemun-gu, Seoul 03722, Korea
Interests: microfluidics; nanotechnology; biochips; biosensors; cell analysis

Special Issue Information

Dear Colleagues,

Circulating biomarkers, which include circulating tumor cells (CTCs), circulating cell-free DNA (cfDNA), extracellular vesicles (exosomes, ectosomes, apoptotic bodies, etc.), and proteins, are a growing research area due to their significance in the diagnosis and prognosis of many diseases; not only cancer, but also other severe symptoms such as metabolic or cardiovascular diseases.

Microfluidic technology also provides a number of useful capabilities for the research of circulating biomarkers: The ability to use very small amounts of samples and reagents, to carry out separations and detections with a high resolution and sensitivity, to reduce the loss of target biomarkers by continuously processing all steps, from sample pre-treatment to analysis, to easily integrate with other techniques, such as electronics and optics, which improve the efficiency of the device, and so on.

Accordingly, this Special Issue will focus on novel microfluidic-based platforms for the isolation, enrichment, and the characterization of circulating biomarkers. Additionally, we would like to discuss advanced approaches for converting experiments at the laboratory scale into clinical practice.

Prof. Dr. Hyo-il Jung
Guest Editor

Manuscript Submission Information

Manuscripts should be submitted online at www.mdpi.com by registering and logging in to this website. Once you are registered, click here to go to the submission form. Manuscripts can be submitted until the deadline. All submissions that pass pre-check are peer-reviewed. Accepted papers will be published continuously in the journal (as soon as accepted) and will be listed together on the special issue website. Research articles, review articles as well as short communications are invited. For planned papers, a title and short abstract (about 100 words) can be sent to the Editorial Office for announcement on this website.

Submitted manuscripts should not have been published previously, nor be under consideration for publication elsewhere (except conference proceedings papers). All manuscripts are thoroughly refereed through a single-blind peer-review process. A guide for authors and other relevant information for submission of manuscripts is available on the Instructions for Authors page. Micromachines is an international peer-reviewed open access monthly journal published by MDPI.

Please visit the Instructions for Authors page before submitting a manuscript. The Article Processing Charge (APC) for publication in this open access journal is 2100 CHF (Swiss Francs). Submitted papers should be well formatted and use good English. Authors may use MDPI's English editing service prior to publication or during author revisions.

Keywords

  • Microfluidics

  • Circulating biomarkers

  • In vitro diagnostics

  • Liquid biopsy

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Published Papers (7 papers)

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Research

Jump to: Review

13 pages, 2681 KiB  
Article
Biometric Image Analysis for Quantitation of Dividing Platelets
by Hyun-Jeong Kim, Yejin Song and Jaewoo Song
Micromachines 2019, 10(1), 1; https://doi.org/10.3390/mi10010001 - 20 Dec 2018
Cited by 13 | Viewed by 3397
Abstract
(1) Background: Quantification of platelet division is challenging because automated Coulter cell counters produce equivocal platelet counts. (2) Methods: We applied the flow cytometric cell tracking dye dilution assay as a popular immunological method to evaluate lymphocyte proliferation to prove and quantitate platelet [...] Read more.
(1) Background: Quantification of platelet division is challenging because automated Coulter cell counters produce equivocal platelet counts. (2) Methods: We applied the flow cytometric cell tracking dye dilution assay as a popular immunological method to evaluate lymphocyte proliferation to prove and quantitate platelet division. We also devised a method relying on platelet culture in a semisolid medium which enabled dividing platelets to be identified by limiting the diffusive movement of platelets. Mixing platelets of different labeling colors in semisolid medium and counting the platelet doublets of each color combination enabled us to prove and quantitate platelet division. (3) Results: The tracking dye dilution assay revealed that 75.5 to 85.6% of platelets were dividing after 20 hours in culture. Platelets labeled with two different tracking dyes were mixed and cultured in semisolid medium for differential doublet counting. We counted platelet singlets and doublets of each color and color combination using confocal microscopy after six hours of culture and compared the relative number of two-colored doublets with binomial prediction to prove platelet division (P < 0.01). Division was suppressed by taxol, nocodazole, or cytochalasin D treatment. We derived a formula for determining the fraction of dividing platelets using the numbers of singlets and doublets of each color and color combination. The platelet division fraction ranged from 8.8 to 17.5%. (4) Conclusion: We successfully measured platelet division using a simple biometric image analysis method with possible future application to microfluidic devices. Full article
(This article belongs to the Special Issue Microfluidics for Circulating Biomarkers)
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15 pages, 2765 KiB  
Article
A Label Free Disposable Device for Rapid Isolation of Rare Tumor Cells from Blood by Ultrasounds
by Itziar González, Julie Earl, Luis J. Fernández, Bruno Sainz, Alberto Pinto, Rosa Monge, Sonia Alcalá, Adela Castillejo, Jose L. Soto and Alfredo Carrato
Micromachines 2018, 9(3), 129; https://doi.org/10.3390/mi9030129 - 15 Mar 2018
Cited by 19 | Viewed by 4858
Abstract
The use of blood samples as liquid biopsy is a label-free method for cancer diagnosis that offers benefits over traditional invasive biopsy techniques. Cell sorting by acoustic waves offers a means to separate rare cells from blood samples based on their physical properties [...] Read more.
The use of blood samples as liquid biopsy is a label-free method for cancer diagnosis that offers benefits over traditional invasive biopsy techniques. Cell sorting by acoustic waves offers a means to separate rare cells from blood samples based on their physical properties in a label-free, contactless and biocompatible manner. Herein, we describe a flow-through separation approach that provides an efficient separation of tumor cells (TCs) from white blood cells (WBCs) in a microfluidic device, “THINUS-Chip” (Thin-Ultrasonic-Separator-Chip), actuated by ultrasounds. We introduce for the first time the concept of plate acoustic waves (PAW) applied to acoustophoresis as a new strategy. It lies in the geometrical chip design: different to other microseparators based on either bulk acoustic waves (BAW) or surface waves (SAW, SSAW and tSAW), it allows the use of polymeric materials without restrictions in the frequency of work. We demonstrate its ability to perform high-throughput isolation of TCs from WBCs, allowing a recovery rate of 84% ± 8% of TCs with a purity higher than 80% and combined viability of 85% at a flow rate of 80 μL/min (4.8 mL/h). The THINUS-Chip performs cell fractionation with low-cost manufacturing processes, opening the door to possible easy printing fabrication. Full article
(This article belongs to the Special Issue Microfluidics for Circulating Biomarkers)
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10 pages, 10420 KiB  
Article
Deterministic Capture of Individual Circulating Tumor Cells Using a Flow-Restricted Microfluidic Trap Array
by Yousang Yoon, Jusin Lee, Ki-Chun Yoo, Onejae Sul, Su-Jae Lee and Seung-Beck Lee
Micromachines 2018, 9(3), 106; https://doi.org/10.3390/mi9030106 - 2 Mar 2018
Cited by 7 | Viewed by 5239
Abstract
Circulating tumor cells (CTCs) are regarded as a strong biomarker which includes clinically valuable information. However, CTCs are very rare and require precise separation and detection for effective clinical applications. Furthermore, downstream analysis has become necessary to identify the distinct sub-population of CTCs [...] Read more.
Circulating tumor cells (CTCs) are regarded as a strong biomarker which includes clinically valuable information. However, CTCs are very rare and require precise separation and detection for effective clinical applications. Furthermore, downstream analysis has become necessary to identify the distinct sub-population of CTCs that causes metastasis. Here, we report a flow-restricted microfluidic trap array capable of deterministic single-cell capture of CTCs. The extent of flow restriction, correlating with the device geometry, was then optimized using a highly invasive breast cancer cell line (LM2 MDA-MB-231) to achieve 97% capture efficiency with a single-cell capture rate of 99%. Single-cell capture of CTCs from mice with full-blown metastasis was also demonstrated. The single-CTC capturing ability of the flow-restricted trap array not only showed cell enumerating ability but also high prospects for application in future automated downstream analysis. Full article
(This article belongs to the Special Issue Microfluidics for Circulating Biomarkers)
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19 pages, 4349 KiB  
Article
The Viability of Single Cancer Cells after Exposure to Hydrodynamic Shear Stresses in a Spiral Microchannel: A Canine Cutaneous Mast Cell Tumor Model
by Dettachai Ketpun, Achariya Sailasuta, Thammawit Suwannaphan, Sudchaya Bhanpattanakul, Alongkorn Pimpin, Werayut Srituravanich, Witsaroot Sripumkhai, Wutthinan Jeamsaksiri and Prapruddee Piyaviriyakul
Micromachines 2018, 9(1), 9; https://doi.org/10.3390/mi9010009 - 28 Dec 2017
Cited by 10 | Viewed by 5737
Abstract
Our laboratory has the fundamental responsibility to study cancer stem cells (CSC) in various models of human and animal neoplasms. However, the major impediments that spike our accomplishment are the lack of universal biomarkers and cellular heterogeneity. To cope with these restrictions, we [...] Read more.
Our laboratory has the fundamental responsibility to study cancer stem cells (CSC) in various models of human and animal neoplasms. However, the major impediments that spike our accomplishment are the lack of universal biomarkers and cellular heterogeneity. To cope with these restrictions, we have tried to apply the concept of single cell analysis, which has hitherto been recommended throughout the world as an imperative solution pack for resolving such dilemmas. Accordingly, our first step was to utilize a predesigned spiral microchannel fabricated by our laboratory to perform size-based single cell separation using mast cell tumor (MCT) cells as a model. However, the impact of hydrodynamic shear stresses (HSS) on mechanical cell injury and viability in a spiral microchannel has not been fully investigated so far. Intuitively, our computational fluid dynamics (CFD) simulation has strongly revealed the formations of fluid shear stress (FSS) and extensional fluid stress (EFS) in the sorting system. The panel of biomedical assays has also disclosed cell degeneration and necrosis in the model. Therefore, we have herein reported the combinatorically detrimental effect of FSS and EFS on the viability of MCT cells after sorting in our spiral microchannel, with discussion on the possibly pathogenic mechanisms of HSS-induced cell injury in the study model. Full article
(This article belongs to the Special Issue Microfluidics for Circulating Biomarkers)
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Review

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21 pages, 2511 KiB  
Review
Progress in Circulating Tumor Cell Research Using Microfluidic Devices
by Hogyeong Gwak, Junmoo Kim, Leila Kashefi-Kheyrabadi, Bongseop Kwak, Kyung-A Hyun and Hyo-Il Jung
Micromachines 2018, 9(7), 353; https://doi.org/10.3390/mi9070353 - 14 Jul 2018
Cited by 39 | Viewed by 7881
Abstract
Circulating tumor cells (CTCs) are a popular topic in cancer research because they can be obtained by liquid biopsy, a minimally invasive procedure with more sample accessibility than tissue biopsy, to monitor a patient’s condition. Over the past decades, CTC research has covered [...] Read more.
Circulating tumor cells (CTCs) are a popular topic in cancer research because they can be obtained by liquid biopsy, a minimally invasive procedure with more sample accessibility than tissue biopsy, to monitor a patient’s condition. Over the past decades, CTC research has covered a wide variety of topics such as enumeration, profiling, and correlation between CTC number and patient overall survival. It is important to isolate and enrich CTCs before performing CTC analysis because CTCs in the blood stream are very rare (0–10 CTCs/mL of blood). Among the various approaches to separating CTCs, here, we review the research trends in the isolation and analysis of CTCs using microfluidics. Microfluidics provides many attractive advantages for CTC studies such as continuous sample processing to reduce target cell loss and easy integration of various functions into a chip, making “do-everything-on-a-chip” possible. However, tumor cells obtained from different sites within a tumor exhibit heterogenetic features. Thus, heterogeneous CTC profiling should be conducted at a single-cell level after isolation to guide the optimal therapeutic path. We describe the studies on single-CTC analysis based on microfluidic devices. Additionally, as a critical concern in CTC studies, we explain the use of CTCs in cancer research, despite their rarity and heterogeneity, compared with other currently emerging circulating biomarkers, including exosomes and cell-free DNA (cfDNA). Finally, the commercialization of products for CTC separation and analysis is discussed. Full article
(This article belongs to the Special Issue Microfluidics for Circulating Biomarkers)
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13 pages, 1711 KiB  
Review
Salivary Exosome and Cell-Free DNA for Cancer Detection
by Kyung-A Hyun, Hogyeong Gwak, Jaehun Lee, Bongseop Kwak and Hyo-Il Jung
Micromachines 2018, 9(7), 340; https://doi.org/10.3390/mi9070340 - 4 Jul 2018
Cited by 50 | Viewed by 10368
Abstract
Liquid biopsies are easier to acquire patient derived samples than conventional tissue biopsies, and their use enables real-time monitoring of the disease through continuous sampling after initial diagnosis, resulting in a paradigm shift to customized treatment according to the patient’s prognosis. Among the [...] Read more.
Liquid biopsies are easier to acquire patient derived samples than conventional tissue biopsies, and their use enables real-time monitoring of the disease through continuous sampling after initial diagnosis, resulting in a paradigm shift to customized treatment according to the patient’s prognosis. Among the various liquid biopsy samples, saliva is easily obtained by spitting or swab sucking without needing an expert for sample collection. In addition, it is known that disease related biomarkers that exist in the blood and have undergone extensive research exist in saliva even at a lower concentration than the blood. Thus, interest in the use of saliva as a liquid biopsy has increased. In this review, we focused on the salivary exosome and cell-free DNA (cfDNA) among the various biomarkers in saliva. Since the exosome and cfDNA in saliva are present at lower concentrations than the biomarkers in blood, it is important to separate and concentrate them before conducting down-stream analyses such as exosome cargo analysis, quantitative polymerase chain reaction (qPCR), and sequencing. However, saliva is difficult to apply directly to microfluidics-based systems for separation because of its high viscosity and the presence of various foreign substances. Therefore, we reviewed the microfluidics-based saliva pretreatment method and then compared the commercially available kit and the microfluidic chip for isolation and enrichment of the exosome and cfDNA in saliva. Full article
(This article belongs to the Special Issue Microfluidics for Circulating Biomarkers)
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18 pages, 2058 KiB  
Review
Liquid Biopsy in Lung Cancer: Clinical Applications of Circulating Biomarkers (CTCs and ctDNA)
by Minji Lim, Chi-Ju Kim, Vijaya Sunkara, Mi-Hyun Kim and Yoon-Kyoung Cho
Micromachines 2018, 9(3), 100; https://doi.org/10.3390/mi9030100 - 28 Feb 2018
Cited by 77 | Viewed by 12805
Abstract
Lung cancer is by far the leading cause of cancer death worldwide, with non-small cell lung cancer (NSCLC) accounting for the majority of cases. Recent advances in the understanding of the biology of tumors and in highly sensitive detection technologies for molecular analysis [...] Read more.
Lung cancer is by far the leading cause of cancer death worldwide, with non-small cell lung cancer (NSCLC) accounting for the majority of cases. Recent advances in the understanding of the biology of tumors and in highly sensitive detection technologies for molecular analysis offer targeted therapies, such as epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors. However, our understanding of an individual patient’s lung cancer is often limited by tumor accessibility because of the high risk and invasive nature of current tissue biopsy procedures. “Liquid biopsy”, the analysis of circulating biomarkers from peripheral blood, such as circulating tumor cells (CTCs) and circulating tumor DNA (ctDNA), offers a new source of cancer-derived materials that may reflect the status of the disease better and thereby contribute to more personalized treatment. In this review, we examined the clinical significance and uniqueness of CTCs and ctDNA from NSCLC patients, isolation and detection methods developed to analyze each type of circulating biomarker, and examples of clinical studies of potential applications for early diagnosis, prognosis, treatment monitoring, and prediction of resistance to therapy. We also discuss challenges that remain to be addressed before such tools are implemented for routine use in clinical settings. Full article
(This article belongs to the Special Issue Microfluidics for Circulating Biomarkers)
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