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 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.

Fabrication and Testing of a Magnetophoretic Bioseparation Chip for Isolation and Detection of Circulating Tumor Cells From Peripheral Blood

2019 ◽  
Author(s):  
Seth Jackson ◽  
Jeff Darabi ◽  
Joseph Schober
Keyword(s):  
Blood Sample ◽  
Cancer Cells ◽  
Tumor Cells ◽  
Circulating Tumor Cells ◽  
Peripheral Blood ◽  
Point Of Care ◽  
Magnetic Field Gradient ◽  
The Body ◽  
Target Cells ◽  
Economic Point

Abstract Significant research involving the isolation and detection of circulating tumor cells (CTCs) has become prevalent in the field of biomedicine. It plays a crucial role in the diagnosis and treatment of cancer and has made substantial strides in recent years. A major event in the timeline of cancer is metastasis, a set of occurrences where cells are shed from a cancerous site, then flow through the circulatory system and seed themselves throughout the body, forming secondary tumors. There are few observable symptoms in the early stages of metastasis and this fact severely limits clinical treatment. The fabrication and preliminary testing of a magnetophoretic bioseparation chip capable of isolating and detecting CTCs from peripheral blood, which can aid in early detection of metastases, is presented in this work. MCF7 breast cancer cells along with superparamagnetic microparticles, which are specifically coated with anti-EpCAM to bind to the cancer cells, are spiked into a blood sample. After the spiked blood sample is introduced into the biochip, a locally engineered magnetic field gradient captures the magnetically labeled cancer cells while the non-target cells are allowed to pass by. Once the target cells are isolated from the blood sample, flow cytometry is used to determine the recovery rate of the magnetophoretic device. The proposed device can operate at continuous flow rates magnitudes higher than other CTC isolation devices and is fabricated using much simpler methods which make it quite unique. These properties combined with greater than 80% recovery rates make the device quite favorable for economic point of care use in clinical applications.

scholarly journals Micro-Raman of Cancer Cells: Toward Label-Free Sorting of Circulating Tumor Cells from Whole Blood

2012 ◽  
Vol 102 (3) ◽  
pp. 589a
Author(s):  
Gregory L. Futia ◽  
Allison G. Caster ◽  
Omid Masihzadeh ◽  
Kian Behbakht ◽  
Emily A. Gibson
Keyword(s):  
Cancer Cells ◽  
Tumor Cells ◽  
Circulating Tumor Cells ◽  
Whole Blood ◽  
Label Free

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.

Concordancy of immunocytochemistry profiling of circulating tumor cells with immunohistochemistry for analysis of therapeutically relevant biomarkers.

2021 ◽  
Vol 39 (15_suppl) ◽  
pp. 3047-3047
Author(s):  
Dadasaheb B Akolkar ◽  
Darshana Patil ◽  
Sudha S Murthy ◽  
Kiran Bendale ◽  
Revati Patil ◽  
...  
Keyword(s):  
Tumor Cells ◽  
Circulating Tumor Cells ◽  
Peripheral Blood ◽  
Tumor Tissue ◽  
Checkpoint Inhibitor ◽  
Matched Pairs ◽  
Tissue Analysis ◽  
The Status ◽  
Checkpoint Inhibitor Therapy ◽  
Selection Of

3047 Background: Immunohistochemistry (IHC) profiling of tumor tissue is the present standard for evaluation of therapeutically relevant biomarkers such as ER, PR, HER2, AR, ARv7, PD-L1 and MMR for selection of targeted, endocrine and checkpoint inhibitor therapy selection. However, this critical analysis is dependent on availability of tumor tissue obtained by an invasive biopsy. Challenges to this analysis include insufficient tumor tissue and inability to perform a repeat biopsy to obtain fresh tumor tissue. We have previously described an approach for negative enrichment of Circulating Tumor Cells (CTCs) from peripheral blood and for Immunocytochemistry (ICC) profiling of these CTCs for detection of diagnostically relevant tissue of origin and subtype specific markers, concordant with tumor tissue analysis. Methods: In the present study, we determined concordance between tumor tissue (HPE) and CTCs (ICC) for ascertaining the status of therapeutically relevant markers ER, PR, HER2, AR, ARv7 PD-L1 and MMR. We evaluated 201 matched pairs of tumor tissue (FFPE blocks) and CTCs obtained from peripheral blood. Results: Among the 743 paired assays on matched tumor tissue and CTCs, concordance (positive or negative) was observed in 651 matched pairs (87.6%). The concordance was 82.9% for ER, 100% for PR, 90.2 % for Her2, 93.8% for AR, 90% for Arv7, 85.1% and 87.6% for PD-L1 clones 22c3 and 28-8, and 85.6% for MMR (MLH1, MSH2, MSH6, and PMS2). Conclusions: The study findings indicate that ICC analysis of CTCs may be able to substitute IHC analysis of tumor tissue for profiling of therapeutically relevant markers. This approach may have application in cases where tumor tissue may be limiting and / or where an invasive biopsy to obtain tumor tissue may be unviable.

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.

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