Cytopathologic identification of circulating tumor cells (CTCs) in breast cancer: Application of size-based enrichment

Kamal, Mohamed; Leslie, Macall; Horton, Crista; Hills, Natalie; Davis, Rebecca; Nguyen, Richard D.; Razaq, Mohammad; Moxley, Katherine M.; Hofman, Paul; Zhang, Roy; Tanaka, Takemi

doi:10.15761/cdp.1000136

Cited by 4 publications

(6 citation statements)

References 38 publications

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“…Studies reported the feasibility of detection of CTCs using isolation by size-based Epithelial/Trophoblastic Tumor cells (ISET ® ) filters and stain by May–Grünwald–Giemsa in conjunction with identification criteria of nuclear irregularity, negative reactivity to immune cell marker as well as endothelial cell markers, and presentation of visible cytoplasm [ 26 ]. To test the negativity of CTC for CD31 in IF×3, we used the additional staining kit for DAPI/CK-FITC/CD31-PE/CD45-Cy5.…”

Section: Discussionmentioning

confidence: 99%

“…Our parallel double-detection format for CTC determination is efficient as it can be ready for pathological evaluation within the standard working hours of one day. The procedure is laboratory friendly and requires basic equipment and microscopes, and can be carried out with a standard grad-school laboratory setup compared with the FDA-approved CellSearch semi-automated CTC detection system or the CTC detection sensitivity of ISET [ 26 ] or using an immunomagnetic enrichment [ 25 ]. Hence, the method is cost-effective, and the cost of the consumables per 7.5 mL blood sample can be estimated at around $500 only.…”

Section: Discussionmentioning

confidence: 99%

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A Laboratory-Friendly CTC Identification: Comparable Double-Immunocytochemistry with Triple-Immunofluorescence

Sulaiman

Aske

et al. 2022

Cancers

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The source of circulating tumor cells (CTC) in the peripheral blood of patients with solid tumors are from primary cancer, metastatic sites, and a disseminated tumor cell pool. As 90% of cancer-related deaths are caused by metastatic progression and/or resistance-associated treatment failure, the above fact justifies the undeniable predictive and prognostic value of identifying CTC in the bloodstream at stages of the disease progression and resistance to treatment. Yet enumeration of CTC remains far from a standard routine procedure either for post-surgery follow-ups or ongoing adjuvant therapy. The most compelling explanation for this paradox is the absence of a convenient, laboratory-friendly, and cost-effective method to determine CTC. We presented a specific and sensitive laboratory-friendly parallel double-detection format method for the simultaneous isolation and identification of CTC from peripheral blood of 91 consented and enrolled patients with various malignant solid tumors of the lung, endometrium, ovary, esophagus, prostate, and liver. Using a pressure-guided method, we used the size-based isolation to capture CTC on a commercially available microfilter. CTC identification was carried out by two expression marker-based independent staining methods, double-immunocytochemistry parallel to standard triple-immunofluorescence. The choice of markers included specific markers for epithelial cells, EpCAM and CK8,18,19, and exclusion markers for WBC, CD45. We tested the method’s specificity based on the validation of the staining method, which included positive and negative spiked samples, blood from the healthy age-matched donor, healthy age-matched leucopaks, and blood from metastatic patients. Our user-friendly cost-effective CTC detection technique may facilitate the regular use of CTC detection even in community-based cancer centers for prognosis, before and after surgery.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

A Laboratory-Friendly CTC Identification: Comparable Double-Immunocytochemistry with Triple-Immunofluorescence

Sulaiman

Aske

et al. 2022

Cancers

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“…Also, this group also used immunofluorescent assay using immunofluorescent microscopy using A45-B/B3 and CD45 antibodies. Another method recently developed by Kamal et al utilizes a size-based exclusion principle for CTC enrichment which allows more sensitive capture of CTCs with minimal biological bias [31].…”

Section: Enrichment Isolation and Enumeration Of Circulating Tumor Cellsmentioning

confidence: 99%

Evaluating the Potential for Circulating Tumor Cells as a Diagnostic and Prognostic Biomarker in Various Cancer Types

Horton¹,

Razaq²

2020

COCB

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Circulating tumor cells (CTCs) are cells that can be found circulating in the peripheral blood of cancer patients. They originate from primary solid tumors and are thought to contribute to metastases and poor prognosis. Since cancer treatment is shifting toward greater personalization, a major goal in the field is the development of less invasive and more cost-effective measures in diagnosis, staging, treatment, prognostic implications, and surveillance of cancer. Utilizing CTCs as a biomarker from a “liquid biopsy” or sample of patients’ blood would be transformative in accomplishing this goal. In this review, we aim to critically assess current pre-clinical and clinical literature over the past two decades implicating CTCs’ potential for use as a predictive biomarker in various cancer types either in addition to or instead of current standards of care. We also are interested in understanding several aspects of CTCs including the role CTCs play in resistance to treatment, the immune system evasion properties of CTCs, the feasibility of using CTCs in clinical practice, and the utility of CTCs for predicting outcomes including patient survival. Furthermore, here we discuss gaps in the literature, limitations of CTCs, potential for their other uses, as well as the significance of CTC detection in patients following surgery.

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Section: Introductionmentioning

confidence: 99%

“…11 Probably for these reasons, data regarding basic CTC morphology in breast cancer are limited and thus the selection of the appropriate separation method is of outmost importance for proper cytopathological identification. [11][12][13][14] There are few published data how separation techniques affect CTCs morphology. [13][14][15][16] Therefore, we designed a study aimed to select the separation method that would allow the identification of CTCs based on their morphology that could be integrated in a routine cytopathology laboratory.…”

Section: Introductionmentioning

confidence: 99%

Morphological features of breast cancer circulating tumor cells in blood after physical and biological type of isolation

Jesenko

Modic

Kuhar

et al. 2021

Radiology and Oncology

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Background Circulating tumor cells (CTCs) have become an important biomarker in breast cancer. Different isolation tech-niques based on their biological or physical features were established. Currently, the most widely used methods for visualization after their separation are based on immunofluorescent staining, which does not provide the information on the morphology. Materials and methods The aim of this study was to evaluate how two different separation techniques affect cell morphology and to analyse cell morphology with techniques used in routine cytopathological laboratory. A direct side-by-side comparison of physical (Parsortix®) and biological (MACS®) separation technique was performed. Results In the preclinical setting, both isolation techniques retained the viability and antigenic characteristics of MCF7 breast cancer cells. Some signs of degeneration such as cell swelling, cytoplasmic blebs, villous projections and vacuolization were observed. In metastatic breast cancer patient cohort, morphological features of isolated CTCs were dependent on the separation technique. After physical separation, CTCs with preserved cell morphology were detected. After biological separation the majority of the isolated CTCs were so degenerated that their identity was difficult to confirm. Conclusions Taken together, physical separation is a suitable technique for detection of CTCs with preserved cell morphology for the use in a routine cytopathological laboratory.

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Cytopathologic identification of circulating tumor cells (CTCs) in breast cancer: Application of size-based enrichment

Cited by 4 publications

References 38 publications

A Laboratory-Friendly CTC Identification: Comparable Double-Immunocytochemistry with Triple-Immunofluorescence

A Laboratory-Friendly CTC Identification: Comparable Double-Immunocytochemistry with Triple-Immunofluorescence

Evaluating the Potential for Circulating Tumor Cells as a Diagnostic and Prognostic Biomarker in Various Cancer Types

Morphological features of breast cancer circulating tumor cells in blood after physical and biological type of isolation

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