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.

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.

Direct freezing of whole blood enables analysis of leucocyte markers by flow cytometry: a proof-of-concept study

2021 ◽  
Author(s):  
Pénélope Bourgoin ◽  
Inès Ait Belkacem ◽  
Isabelle Arnoux ◽  
Pierre-Emmanuel Morange ◽  
Fabrice Malergue
Keyword(s):  
Flow Cytometry ◽  
Whole Blood ◽  
Blood Cells ◽  
White Blood Cells ◽  
Proof Of Concept ◽  
Fresh Blood ◽  
Blood Samples ◽  
Step Flow ◽  
Freezing Method ◽  
One Step

Aim: A new one-step flow cytometry procedure has been recently demonstrated for identifying subjects with infections, but only for fresh whole blood samples. The goal of this study was to assess its applicability on frozen samples, by proposing a new method to perform the sample freezing directly and easily. Methods: Fresh blood was tested, then frozen either directly or with dimethylsulfoxide and serum. Common markers of white blood cells as well as infection-related biomarkers were tested. Results: All percentages of leucocyte subsets and levels of infection-related biomarkers were significantly correlated between frozen and fresh samples. Conclusion: The direct freezing method enables an accurate assessment of common cellular sub-populations and of levels of important infectious biomarkers via flow cytometry.

scholarly journals Flexible Micro Spring Array Device for High-Throughput Enrichment of Viable Circulating Tumor Cells

2014 ◽  
Vol 60 (2) ◽  
pp. 323-333 ◽  
Author(s):  
Ramdane A Harouaka ◽  
Ming-Da Zhou ◽  
Yin-Ting Yeh ◽  
Waleed J Khan ◽  
Avisnata Das ◽  
...  
Keyword(s):  
Tumor Cells ◽  
Circulating Tumor Cells ◽  
Whole Blood ◽  
Ex Vivo ◽  
Cell Damage ◽  
Flexible Structures ◽  
Capture Efficiency ◽  
Clinical Samples ◽  
Blood Samples ◽  
Whole Blood Samples

Abstract BACKGROUND The dissemination of circulating tumor cells (CTCs) that cause metastases in distant organs accounts for the majority of cancer-related deaths. CTCs have been established as a cancer biomarker of known prognostic value. The enrichment of viable CTCs for ex vivo analysis could further improve cancer diagnosis and guide treatment selection. We designed a new flexible micro spring array (FMSA) device for the enrichment of viable CTCs independent of antigen expression. METHODS Unlike previous microfiltration devices, flexible structures at the micro scale minimize cell damage to preserve viability, while maximizing throughput to allow rapid enrichment directly from whole blood with no need for sample preprocessing. Device performance with respect to capture efficiency, enrichment against leukocytes, viability, and proliferability was characterized. CTCs and CTC microclusters were enriched from clinical samples obtained from breast, lung, and colorectal cancer patients. RESULTS The FMSA device enriched tumor cells with 90% capture efficiency, higher than 104 enrichment, and better than 80% viability from 7.5-mL whole blood samples in <10 min on a 0.5-cm2 device. The FMSA detected at least 1 CTC in 16 out of 21 clinical samples (approximately 76%) compared to 4 out of 18 (approximately 22%) detected with the commercial CellSearch® system. There was no incidence of clogging in over 100 tested fresh whole blood samples. CONCLUSIONS The FMSA device provides a versatile platform capable of viable enrichment and analysis of CTCs from clinically relevant volumes of whole blood.

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.

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.

scholarly journals Detection of PD-L1 in circulating tumor cells and white blood cells from patients with advanced non-small-cell lung cancer

2018 ◽  
Vol 29 (1) ◽  
pp. 193-199 ◽  
Author(s):  
M. Ilié ◽  
E. Szafer-Glusman ◽  
V. Hofman ◽  
E. Chamorey ◽  
S. Lalvée ◽  
...  
Keyword(s):  
Lung Cancer ◽  
Small Cell Lung Cancer ◽  
Tumor Cells ◽  
Circulating Tumor Cells ◽  
Cell Lung Cancer ◽  
Blood Cells ◽  
White Blood Cells ◽  
Small Cell ◽  
Small Cell Lung
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