Enhanced Luminescent Detection of Circulating Tumor Cells by a 3D Printed Immunomagnetic Concentrator

Circulating tumor cells (CTCs) are an indicator of metastatic progression and relapse. Since non-CTC cells such as red blood cells outnumber CTCs in the blood, the separation and enrichment of CTCs is key to improving their detection sensitivity. The ATP luminescence assay can measure intracellular ATP to detect cells quickly but has not yet been used for CTC detection in the blood because extracellular ATP in the blood, derived from non-CTCs, interferes with the measurement. Herein, we report on the improvement of the ATP luminescence assay for the detection of CTCs by separating and concentrating CTCs in the blood using a 3D printed immunomagnetic concentrator (3DPIC). Because of its high-aspect-ratio structure and resistance to high flow rates, 3DPIC allows cancer cells in 10 mL to be concentrated 100 times within minutes. This enables the ATP luminescence assay to detect as low as 10 cells in blood, thereby being about 10 times more sensitive than when commercial kits are used for CTC concentration. This is the first time that the ATP luminescence assay was used for the detection of cancer cells in blood. These results demonstrate the feasibility of 3DPIC as a concentrator to improve the detection limit of the ATP luminescence assay for the detection of CTCs.

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Positively Charged Magnetic Nanoparticles for Capture of Circulating Tumor Cells from Clinical Blood Samples

Nano LIFE ◽

10.1142/s1793984419710016 ◽

2020 ◽

Vol 10 (03) ◽

pp. 1971001 ◽

Cited By ~ 1

Author(s):

Shengming Wu ◽

Yilong Wang ◽

Donglu Shi

Keyword(s):

Magnetic Nanoparticles ◽

Cancer Cells ◽

Cancer Patients ◽

Tumor Cells ◽

Circulating Tumor Cells ◽

Cancer Cell ◽

Detection Sensitivity ◽

Blood Samples ◽

Charged Nanoparticles ◽

Positively Charged

Isolation of circulating tumor cells (CTCs) from cancer patients is of high value for disease monitoring and metastasis diagnosis. Although many new detection methods have emerged in recent years, the detection of CTCs is a current challenge due to lack of specific and sensitive markers. In our previous work, cancer cell surfaces, from over 20 cancer cell lines, have been shown to be negatively-charged regardless of their phenotype by using electrically-charged nanoparticles as a probe. The strong electrostatic interaction between the negative cancer cells and positively charged nanoparticles can well remain in a physiological liquid environment in the presence of serum proteins, enabling effective binding between them. As a result, the cancer cells can be magnetically separated by employing an external magnet. In this technical report, we present preliminary results on the investigation of CTC isolation from both mimetic and clinical blood samples. We show high CTC detection sensitivity by the positively-charged magnetic nanoparticles (PMNs) even at the original concentration of 10 cells per mL mimetic blood sample. The CTCs in the peripheral blood of colorectal cancer patients were isolated and identified by cellular morphology and immunofluorescence staining.

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The Mechanical Fingerprint of Circulating Tumor Cells (CTCs) in Breast Cancer Patients

Cancers ◽

10.3390/cancers13051119 ◽

2021 ◽

Vol 13 (5) ◽

pp. 1119

Author(s):

Ivonne Nel ◽

Erik W. Morawetz ◽

Dimitrij Tschodu ◽

Josef A. Käs ◽

Bahriye Aktas

Keyword(s):

Breast Cancer ◽

Cancer Cells ◽

Cancer Patients ◽

Tumor Cells ◽

Circulating Tumor Cells ◽

Hematopoietic Cells ◽

Surrogate Marker ◽

Clinical Samples ◽

Breast Cancer Patients ◽

Shape Restoration

Circulating tumor cells (CTCs) are a potential predictive surrogate marker for disease monitoring. Due to the sparse knowledge about their phenotype and its changes during cancer progression and treatment response, CTC isolation remains challenging. Here we focused on the mechanical characterization of circulating non-hematopoietic cells from breast cancer patients to evaluate its utility for CTC detection. For proof of premise, we used healthy peripheral blood mononuclear cells (PBMCs), human MDA-MB 231 breast cancer cells and human HL-60 leukemia cells to create a CTC model system. For translational experiments CD45 negative cells—possible CTCs—were isolated from blood samples of patients with mamma carcinoma. Cells were mechanically characterized in the optical stretcher (OS). Active and passive cell mechanical data were related with physiological descriptors by a random forest (RF) classifier to identify cell type specific properties. Cancer cells were well distinguishable from PBMC in cell line tests. Analysis of clinical samples revealed that in PBMC the elliptic deformation was significantly increased compared to non-hematopoietic cells. Interestingly, non-hematopoietic cells showed significantly higher shape restoration. Based on Kelvin–Voigt modeling, the RF algorithm revealed that elliptic deformation and shape restoration were crucial parameters and that the OS discriminated non-hematopoietic cells from PBMC with an accuracy of 0.69, a sensitivity of 0.74, and specificity of 0.63. The CD45 negative cell population in the blood of breast cancer patients is mechanically distinguishable from healthy PBMC. Together with cell morphology, the mechanical fingerprint might be an appropriate tool for marker-free CTC detection.

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Circulating Tumor Cells from Enumeration to Analysis: Current Challenges and Future Opportunities

Cancers ◽

10.3390/cancers13112723 ◽

2021 ◽

Vol 13 (11) ◽

pp. 2723

Author(s):

Yu-Ping Yang ◽

Teresa M. Giret ◽

Richard J. Cote

Keyword(s):

Tumor Cells ◽

Circulating Tumor Cells ◽

Ex Vivo ◽

Detection Sensitivity ◽

Clinical Utilization ◽

Diagnosis And Prognosis ◽

Early Cancer Diagnosis ◽

Potential Marker ◽

Potential Applications ◽

Downstream Analysis

Circulating tumor cells (CTCs) have been recognized as a major contributor to distant metastasis. Their unique role as metastatic seeds renders them a potential marker in the circulation for early cancer diagnosis and prognosis as well as monitoring of therapeutic response. In the past decade, researchers mainly focused on the development of isolation techniques for improving the recovery rate and purity of CTCs. These developed techniques have significantly increased the detection sensitivity and enumeration accuracy of CTCs. Currently, significant efforts have been made toward comprehensive molecular characterization, ex vivo expansion of CTCs, and understanding the interactions between CTCs and their associated cells (e.g., immune cells and stromal cells) in the circulation. In this review, we briefly summarize existing CTC isolation technologies and specifically focus on advances in downstream analysis of CTCs and their potential applications in precision medicine. We also discuss the current challenges and future opportunities in their clinical utilization.

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

10.1115/fedsm2021-65766 ◽

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.

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Circulating Tumor Cells: Diagnostic and Therapeutic Applications

Annual Review of Biomedical Engineering ◽

10.1146/annurev-bioeng-062117-120947 ◽

2018 ◽

Vol 20 (1) ◽

pp. 329-352 ◽

Cited By ~ 27

Author(s):

Eric Lin ◽

Thong Cao ◽

Sunitha Nagrath ◽

Michael R. King

Keyword(s):

Cancer Cells ◽

Tumor Cells ◽

Circulating Tumor Cells ◽

Poor Prognosis ◽

Experimental Therapeutics ◽

Next Generation ◽

Therapeutic Applications

Metastasis contributes to poor prognosis in many types of cancer and is the leading cause of cancer-related deaths. Tumor cells metastasize to distant sites via the circulatory and lymphatic systems. In this review, we discuss the potential of circulating tumor cells for diagnosis and describe the experimental therapeutics that aim to target these disseminating cancer cells. We discuss the advantages and limitations of such strategies and how they may lead to the development of the next generation of antimetastasis treatments.

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The combination of size-based separation and selection-free technology provides higher circulating tumor cells detection sensitivity than either method alone in patients with metastatic prostate cancer

BJU International ◽

10.1111/bju.15041 ◽

2020 ◽

Cited By ~ 3

Author(s):

Liang Dong ◽

Zhongyuan Zhang ◽

Kimberly Smith ◽

Morgan Kuczler ◽

Diane Reyes ◽

...

Keyword(s):

Prostate Cancer ◽

Tumor Cells ◽

Circulating Tumor Cells ◽

Metastatic Prostate Cancer ◽

Detection Sensitivity ◽

Size Based Separation

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

Lab on a Chip ◽

10.1039/c9lc00575g ◽

2019 ◽

Vol 19 (20) ◽

pp. 3427-3437 ◽

Cited By ~ 6

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.

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Nanotechnology-Assisted Isolation and Analysis of Circulating Tumor Cells on Microfluidic Devices

Micromachines ◽

10.3390/mi11080774 ◽

2020 ◽

Vol 11 (8) ◽

pp. 774 ◽

Cited By ~ 2

Author(s):

Jie Cheng ◽

Yang Liu ◽

Yang Zhao ◽

Lina Zhang ◽

Lingqian Zhang ◽

...

Keyword(s):

Tumor Cells ◽

Circulating Tumor Cells ◽

Liquid Biopsy ◽

Cancer Metastasis ◽

Human Peripheral Blood ◽

Microfluidic Devices ◽

Detection Sensitivity ◽

Cell Capture ◽

Primary Tumors ◽

Downstream Analysis

Circulating tumor cells (CTCs), a type of cancer cell that spreads from primary tumors into human peripheral blood and are considered as a new biomarker of cancer liquid biopsy. It provides the direction for understanding the biology of cancer metastasis and progression. Isolation and analysis of CTCs offer the possibility for early cancer detection and dynamic prognosis monitoring. The extremely low quantity and high heterogeneity of CTCs are the major challenges for the application of CTCs in liquid biopsy. There have been significant research endeavors to develop efficient and reliable approaches to CTC isolation and analysis in the past few decades. With the advancement of microfabrication and nanomaterials, a variety of approaches have now emerged for CTC isolation and analysis on microfluidic platforms combined with nanotechnology. These new approaches show advantages in terms of cell capture efficiency, purity, detection sensitivity and specificity. This review focuses on recent progress in the field of nanotechnology-assisted microfluidics for CTC isolation and detection. Firstly, CTC isolation approaches using nanomaterial-based microfluidic devices are summarized and discussed. The different strategies for CTC release from the devices are specifically outlined. In addition, existing nanotechnology-assisted methods for CTC downstream analysis are summarized. Some perspectives are discussed on the challenges of current methods for CTC studies and promising research directions.

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