scholarly journals Enrichment of Circulating Tumor Cells from Whole Blood Using a Microfluidic Device for Sequential Physical and Magnetophoretic Separations

Micromachines ◽  
2020 ◽  
Vol 11 (5) ◽  
pp. 481 ◽  
Author(s):  
Jusin Lee ◽  
Onejae Sul ◽  
Seung-Beck Lee
Keyword(s):  
Specific Surface ◽  
Cancer Cells ◽  
Tumor Cells ◽  
Circulating Tumor Cells ◽  
Microfluidic Device ◽  
Whole Blood ◽  
Blood Cells ◽  
Magnetic Beads ◽  
Separation Efficiency ◽  
White Blood Cells

Based on their high clinical potential, the isolation and enrichment of rare circulating tumor cells (CTCs) from peripheral blood cells has been widely investigated. There have been technical challenges with CTC separation methods using solely cancer-specific surface molecules or just using physical properties of CTCs, as they may suffer from heterogeneity or lack of specificity from overlapping physical characteristics with leukocytes. Here, we integrated an immunomagnetic-based negative enrichment method that utilizes magnetic beads attached to leukocyte-specific surface antigens, with a physical separation method that utilizes the distinct size and deformability of CTCs. By manipulating the pressure distribution throughout the device and balancing the drag and magnetic forces acting on the magnetically labeled white blood cells (WBCs), the sequential physical and magnetophoretic separations were optimized to isolate intact cancer cells, regardless of heterogeneity from whole blood. Using a breast cancer cell line in whole blood, we achieved 100% separation efficiency for cancer cells and an average of 97.2% for WBCs, which resulted in a 93.3% average separation purity. The experimental results demonstrated that our microfluidic device can be a promising candidate for liquid biopsy and can be a vital tool for aiding future cancer research.

scholarly journals Continuous Separation of Circulating Tumor Cells from Whole Blood Using a Slanted Weir Microfluidic Device

Cancers ◽  
2019 ◽  
Vol 11 (2) ◽  
pp. 200 ◽  
Author(s):  
Yousang Yoon ◽  
Jusin Lee ◽  
Moonsoo Ra ◽  
Hyeokshin Gwon ◽  
Seungwon Lee ◽  
...  
Keyword(s):  
Tumor Cells ◽  
Circulating Tumor Cells ◽  
Microfluidic Device ◽  
Whole Blood ◽  
Tumor Heterogeneity ◽  
Separation Efficiency ◽  
Cancer Cell Line ◽  
Analysis Tool ◽  
Surface Molecules ◽  
Clinical Potential

The separation of circulating tumor cells (CTCs) from the peripheral blood is an important issue that has been highlighted because of their high clinical potential. However, techniques that depend solely on tumor-specific surface molecules or just the larger size of CTCs are limited by tumor heterogeneity. Here, we present a slanted weir microfluidic device that utilizes the size and deformability of CTCs to separate them from the unprocessed whole blood. By testing its ability using a highly invasive breast cancer cell line, our device achieved a 97% separation efficiency, while showing an 8-log depletion of erythrocytes and 5.6-log depletion of leukocytes. We also developed an image analysis tool that was able to characterize the various morphologies and differing deformability of the separating cells. From the results, we believe our system possesses a high potential for liquid biopsy, aiding future cancer research.

scholarly journals Negative enrichment of circulating tumor cells from unmanipulated whole blood with a 3D printed device

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Chia-Heng Chu ◽  
Ruxiu Liu ◽  
Tevhide Ozkaya-Ahmadov ◽  
Brandi E. Swain ◽  
Mert Boya ◽  
...  
Keyword(s):  
Tumor Cells ◽  
Circulating Tumor Cells ◽  
Whole Blood ◽  
Blood Cells ◽  
Point Of Care ◽  
White Blood Cells ◽  
Normal Blood ◽  
Blood Samples ◽  
Membrane Antigens ◽  
Tumor Types

AbstractReliable and routine isolation of circulating tumor cells (CTCs) from peripheral blood would allow effective monitoring of the disease and guide the development of personalized treatments. Negative enrichment of CTCs by depleting normal blood cells ensures against a biased selection of a subpopulation and allows the assay to be applied on different tumor types. Here, we report an additively manufactured microfluidic device that can negatively enrich viable CTCs from clinically-relevant volumes of unmanipulated whole blood samples. Our device depletes nucleated blood cells based on their surface antigens and the smaller anucleated cells based on their size. Enriched CTCs are made available off the device in suspension making our technique compatible with standard immunocytochemical, molecular and functional assays. Our device could achieve a ~ 2.34-log depletion by capturing > 99.5% of white blood cells from 10 mL of whole blood while recovering > 90% of spiked tumor cells. Furthermore, we demonstrated the capability of the device to isolate CTCs from blood samples collected from patients (n = 15) with prostate and pancreatic cancers in a pilot study. A universal CTC assay that can differentiate tumor cells from normal blood cells with the specificity of clinically established membrane antigens yet require no label has the potential to enable routine blood-based tumor biopsies at the point-of-care.

Isolation of Circulating Tumor Cells Using Stiffness-Based Filtration Platform

2015 ◽  
Author(s):  
Emrah Celik ◽  
Nicolas Rongione ◽  
Amelia Bahamonde ◽  
Zheng Ao ◽  
Ram Datar
Keyword(s):  
Tumor Cell ◽  
Cancer Cells ◽  
Tumor Cells ◽  
Circulating Tumor Cells ◽  
Blood Cells ◽  
High Efficiency ◽  
White Blood Cells ◽  
Cell Isolation ◽  
Element Analysis ◽  
Cancer Disease

Analysis of isolated cancer cells in circulation is proven to help determine the success of the cancer treatment and understand the genetic signature of cancer disease. Scarcity of these cells in blood circulation (1–10 CTC in 1ml blood) however, makes the isolation process extremely challenging. Ever improving CTC isolation methods fall into two main categories: 1.Immunomagnetic separation based on antibody binding to tumor specific biomarkers expressed on the cell 2. Physical separation based on the size of the CTCs. Efficiency in cell isolation is still low in these techniques due to the variation in expression level of tumor specific antigens and tumor cell size. Therefore, tumor cell isolation strategies using new CTC biomarkers must be explored. In this study, we investigated the feasibility of using mechanical stiffness difference in order to detect and isolate the circulating tumor cells from the blood cells. AFM nanindentation experiments revealed that cancer cells are significantly softer than the surrounding white blood cells and therefore, stiffness can be used as a biomarker for CTC isolation. In addition, finite element analysis simulations have shown that CTC isolation can be performed at high efficiency using stiffness-based isolation. Therefore, stiffness based isolation has a potential to achieve fast, label-free isolation of CTCs at high efficiency for clinical applications.

A Microfluidic Platform for On-Chip Analysis of Circulating Tumor Cells

2021 ◽  
Author(s):  
Jeff Darabi ◽  
Joseph Schober
Keyword(s):  
Cancer Cells ◽  
Tumor Cells ◽  
Circulating Tumor Cells ◽  
Whole Blood ◽  
Peripheral Blood ◽  
Automated Image Analysis ◽  
Rare Cancer ◽  
On Chip ◽  
Chip Analysis ◽  
Selection Of

Abstract Studies have shown that primary tumor sites begin shedding cancerous cells into peripheral blood at early stages of cancer, and the presence and frequency of circulating tumor cells (CTCs) in blood is directly proportional to disease progression. The challenge is that the concentration of the CTCs in peripheral blood may be extremely low. In the past few years, several microfluidic-based concepts have been investigated to isolate CTCs from whole blood. However, these devices are generally hampered by complex fabrication processes and very low volumetric throughputs, which may not be practical for rapid clinical applications. This paper presents a high-performance yet simple magnetophoretic microfluidic chip for the enrichment and on-chip analysis of rare CTCs from blood. Microscopic and flow cytometric assays developed for selection of cancer cell lines, selection of monoclonal antibodies, and optimization of bead coupling are discussed. Additionally, on-chip characterization of rare cancer cells using high resolution immunofluorescence microscopy and modeling results for prediction of CTC capture length are presented. The device has the ability to interface directly with on-chip pre and post processing modules such as mixing, incubation, and automated image analysis systems. These features will enable us to isolate rare cancer cells from whole blood and detect them on the chip with subcellular resolution.

scholarly journals Hybrid negative enrichment of circulating tumor cells from whole blood in a 3D-printed monolithic device

Lab on a Chip ◽  
2019 ◽  
Vol 19 (20) ◽  
pp. 3427-3437 ◽  
Author(s):  
Chia-Heng Chu ◽  
Ruxiu Liu ◽  
Tevhide Ozkaya-Ahmadov ◽  
Mert Boya ◽  
Brandi E. Swain ◽  
...  
Keyword(s):  
Tumor Cells ◽  
Circulating Tumor Cells ◽  
Microfluidic Device ◽  
Whole Blood ◽  
3D Printed

A monolithic 3D-printed microfluidic device integrated with stacked layers of functionalized leukodepletion channels and microfiltration for the negative enrichment of circulating tumor cells directly from clinically relevant volumes of whole blood.

scholarly journals Chemically modified plastic tube for high volume removal and collection of circulating tumor cells

2015 ◽  
Author(s):  
Angelo Gaitas ◽  
Gwangseong Kim
Keyword(s):  
Cancer Cells ◽  
Tumor Cells ◽  
Circulating Tumor Cells ◽  
Whole Blood ◽  
High Volume ◽  
Blood Stream ◽  
Plastic Tube ◽  
Chemically Modified ◽  
Stage Of Development ◽  
Flow Through

In this preliminary effort, we use a commercially available and chemically modified tube to selectively capture circulating tumor cells (CTCs) from the blood stream by immobilizing human anti-EpCAM antibodies on the tube's interior surface. We describe the steps required to modify a tube into a cancer cell capturing device. Using these simple modifications, at this proof-of-concept stage of development, we were able to capture about 85% of cancer cells from suspension and 44% of cancer cells from spiked whole blood, the capture percentage being dependent on the tube's length and the number of cancer cells present. Previous work by other researchers has focused on extracting small blood volumes and capturing CTCs with complicated micro-fluidic devices for diagnostic purposes. In addition, prior results of other researchers point to a possible reduction in metastasis achieved by removing CTCs from the bloodstream. We believe that with the utilization of appropriate tube lengths and procedures, we can ensure capture and removal of nearly the entire CTC population in whole blood. Following whole blood flow through the tube, the tube can be trypsinized to release the captured live CTCs for further analysis and testing.

Interactions Between Circulating Tumor Cells and Aptamer-Functionalized Microposts in a Flow

2017 ◽  
Author(s):  
Kangfu Chen ◽  
Teodor Georgiev ◽  
Z. Hugh Fan
Keyword(s):  
Tumor Cells ◽  
Circulating Tumor Cells ◽  
Microfluidic Device ◽  
Blood Cells ◽  
Volume Ratio ◽  
Microfluidic Devices ◽  
Cancer Prognosis ◽  
Device Performance ◽  
Simulation Methodology ◽  
Geometry Design

Circulating Tumor Cells (CTCs) have been considered as important biomarkers for cancer prognosis and treatment. However, there are only tens of CTCs in one billion of healthy blood cells. This CTC rarity challenge has been addressed by microfluidics technology that sheds light on efficient CTC detection and isolation. Using antibodies or aptamers to capture CTCs is one of the strategies for CTC isolation. A lot of work has been carried out to improve CTC capture efficiency and purity (i.e., specificity). The main consideration to optimize microfluidic device performance includes increasing surface-area-to-volume ratio and reducing shear stress, both of which are closely related to the interaction between CTCs and the microfluidic device. Here we report a detailed study on the interactions between CTCs and aptamer-functionalized microposts in a microfluidic device. We have evaluated the distribution of captured CTCs around a micropost. In addition, simulation was conducted to model CTC capture patterns around microposts. We found the simulated CTC capture pattern largely agree with the experimental results. The simulation methodology could be applicable for other affinity-based CTC isolation devices and approaches. The goal of the study is to improve the microfluidic device performance and provide a rapid and economical way to optimize the geometry design of the microfluidic devices for CTC isolation.

scholarly journals Fast and Label-Free Isolation of Circulating Tumor Cells from Blood: From a Research Microfluidic Platform to an Automated Fluidic Instrument, VTX-1 Liquid Biopsy System

2018 ◽  
Vol 23 (1) ◽  
pp. 16-29 ◽  
Author(s):  
Clementine A. Lemaire ◽  
Sean Z. Liu ◽  
Charles L. Wilkerson ◽  
Vishnu C. Ramani ◽  
Nasim A. Barzanian ◽  
...  
Keyword(s):  
Tumor Cells ◽  
Circulating Tumor Cells ◽  
Cancer Cell ◽  
Blood Cells ◽  
Liquid Biopsy ◽  
White Blood Cells ◽  
Automated System ◽  
Cell Recovery ◽  
Blood Draw ◽  
Multiple Tumor

Tumor tissue biopsies are invasive, costly, and collect a limited cell population not completely reflective of patient cancer cell diversity. Circulating tumor cells (CTCs) can be isolated from a simple blood draw and may be representative of the diverse biology from multiple tumor sites. The VTX-1 Liquid Biopsy System was designed to automate the isolation of clinically relevant CTC populations, making the CTCs available for easy analysis. We present here the transition from a cutting-edge microfluidic innovation in the lab to a commercial, automated system for isolating CTCs directly from whole blood. As the technology evolved into a commercial system, flexible polydimethylsiloxane microfluidic chips were replaced by rigid poly(methyl methacrylate) chips for a 2.2-fold increase in cell recovery. Automating the fluidic processing with the VTX-1 further improved cancer cell recovery by nearly 1.4-fold, with a 2.8-fold decrease in contaminating white blood cells and overall improved reproducibility. Two isolation protocols were optimized that favor either the cancer cell recovery (up to 71.6% recovery) or sample purity (≤100 white blood cells/mL). The VTX-1’s performance was further tested with three different spiked breast or lung cancer cell lines, with 69.0% to 79.5% cell recovery. Finally, several cancer research applications are presented using the commercial VTX-1 system.

Results of telomerase activity measurements from live circulating tumor cells captured on a slot microfilter in a phase III SWOG-coordinated prostate cancer trial (S0421).

2012 ◽  
Vol 30 (15_suppl) ◽  
pp. 4663-4663
Author(s):  
Amir Goldkorn ◽  
Benjamin Ely ◽  
David I. Quinn ◽  
Catherine M. Tangen ◽  
Yu-Chong Tai ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
Tumor Cells ◽  
Circulating Tumor Cells ◽  
Blood Cells ◽  
White Blood Cells ◽  
Telomerase Activity ◽  
Phase Iii ◽  
Cancer Trial ◽  
Activity Measurements ◽  
Baseline Psa

4663 Background: Analysis of circulating tumor cells (CTC) is a promising biomarker strategy in advanced prostate cancer, and telomerase activity (TA) is a recognized cancer marker. To test whether CTC TA is prognostic for survival (OS), we developed a novel Parylene-C slot microfilter capable of capturing live CTC and used it to measure CTC TA as part of a Phase III SWOG-coordinated therapeutic trial in metastatic castration resistant prostate cancer (S0421). Methods: Blood samples were drawn into EDTA tubes and shipped overnight to a central processing site. After Ficoll centrifugation, low constant pressure was used to pass the mononuclear cell layer through two slot microfilters in series as published previously (filter1 captures CTC + background white blood cells; filter2 captures only background white blood cells). Filter-trapped cells were lysed in CHAPS buffer and assayed for TA using qPCR-based telomeric repeat amplification. In parallel, CTC were enumerated using CellSearch (J&J). Cox regression was used to evaluate the association between baseline (pre-treatment) TA and OS overall, and within subgroups characterized by good prognosis (<5) vs. poor prognosis (>=5) baseline CTC counts. CART regression was used to explore potential prognostic subgroups based on baseline PSA, CTC, and TA cutpoints. Results: Samples were obtained from 263 patients. While no association was observed between TA and OS overall, in patients with baseline CTC >=5 (108 of 263 or 41% of patients), TAfilter2 – TAfilter1 representing high CTC TA relative to background blood cells was associated with a hazard ratio (HR) of 1.14 (95% CI 1.05-1.23, p<0.001) for OS after adjusting for risk factors and remained significant when also adjusting for CTC: HR 1.14 (95% CI 1.04-1.23; p=0.005). Exploratory CART regression assessing baseline PSA, CTC, and TA identified risk groups based only on CTC and TA values. Conclusions: Baseline TA from CTC live-captured on a new slot microfilter is the first CTC biomarker shown to be prognostic of OS in men with CTC counts >=5 in a prospective clinical trial. CTC TA may be useful for further identifying prognostic groups in this population.

Sign in / Sign up

Export Citation Format

Share Document