Effect of Blood Collection Tube Type and Time to Processing on the Enumeration and High-Content Characterization of Circulating Tumor Cells Using the High-Definition Single-Cell Assay

Rodrı́guez-Lee, Mariam; Kolatkar, Anand; McCormick, Madelyn; Dago, Angel D.; Kendall, Jude; Carlsson, Nils Anders; Bethel, Kelly; Greenspan, Emily J.; Hwang, Shelley; Waitman, Kathryn R.; Nieva, Jorgé; Hicks, James; Kühn, Peter

doi:10.5858/arpa.2016-0483-oa

Cited by 44 publications

(54 citation statements)

References 25 publications

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“…Assessing and controlling the preanalytical handling conditions of clinical samples is essential for downstream CTC analysis . Preanalytical variables can lead to in vitro modifications and loss of nucleic acid stability, which both affect the interpretation of results . Several preanalytical issues arise in the interval between PB draw, CTCs enrichment, and their downstream analysis.…”

Section: Quality Control and Preanalytical Parameters In Liquid Biopsiesmentioning

confidence: 99%

“…All these steps are very important for the effective preservation of CTCs, as they can affect the integrity of highly fragile CTCs, rendering their accurate enumeration and characterization hard and ambiguous . Very recently, the effect of blood collection tube types and time to processing on the enumeration and high‐content characterization of CTCs based on imaging was evaluated in detail …”

Section: Quality Control and Preanalytical Parameters In Liquid Biopsiesmentioning

confidence: 99%

“…97 Preanalytical variables can lead to in vitro modifications and loss of nucleic acid stability, which both affect the interpretation of results. 98,99 Several preanalytical issues arise in the interval between PB draw, CTCs enrichment, and their downstream analysis. These include delay in blood processing, storage temperature, and even agitation of the sample during shipment.…”

Section: Quality Control and Preanalytical Parameters In Liquid Biomentioning

confidence: 99%

“…97 Very recently, the effect of blood collection tube types and time to processing on the enumeration and high-content characterization of CTCs based on imaging was evaluated in detail. 99 In liquid biopsy, the development of more advanced applications that rely on detailed molecular analysis of CTCs has been limited by the requirement to process fresh blood within few hours of blood draw. 100 is not specifically designed for CTC analysis, as RNA is stabilized after lysis of all cells present in whole blood.…”

Section: Quality Control and Preanalytical Parameters In Liquid Biomentioning

confidence: 99%

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Liquid biopsies

Lianidou

Pantel

2019

Genes Chromosomes & Cancer

131

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Liquid biopsy is based on minimally invasive blood tests and has a high potential to significantly change the therapeutic strategy in cancer patients, providing an extremely powerful and reliable noninvasive clinical tool for the individual molecular profiling of patients in real time. Liquid biopsy approaches include the analysis of circulating tumor cells (CTCs), circulating tumor DNA (ctDNA), circulating miRNAs, and tumor‐derived extracellular vesicles (EVs) that are shed from primary tumors and their metastatic sites into peripheral blood. The major advantage of liquid biopsy analysis is that it is minimally invasive, and can be serially repeated, thus allowing extracting information from the tumor in real time. Moreover, the identification of predictive biomarkers in peripheral blood that can monitor response to therapy in real time holds a very strong potential for novel approaches in the therapeutic management of cancer patients. In this review, we summarize recent knowledge on CTCs and ctDNA and discuss future trends in the field.

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Section: Quality Control and Preanalytical Parameters In Liquid Biopsiesmentioning

confidence: 99%

Section: Quality Control and Preanalytical Parameters In Liquid Biopsiesmentioning

confidence: 99%

Section: Quality Control and Preanalytical Parameters In Liquid Biomentioning

confidence: 99%

Section: Quality Control and Preanalytical Parameters In Liquid Biomentioning

confidence: 99%

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Liquid biopsies

Lianidou

Pantel

2019

Genes Chromosomes & Cancer

131

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“…Matching blood and tumor touch prep samples were processed in parallel within 24 hours after collection as previously described for peripheral blood [9], with nucleated cells plated in a monolayer of 3*10 6 cells/ slide (Marienfeld, Lauda, Germany) before cryobanking. Slides were stained in batches of 50, using the validated, standard HD-SCA protocol with a cocktail of pancytokeratin (CK), CK-19, CD45 antibodies and DAPI [9,31] complemented with a CDX2 monoclonal antibody EPR2764Y (Abcam, Cambridge, UK), and imaged in 2305 frames per slide using automated high-throughput fluorescence scanning microscopy at 10X magnification with exposures and gain set to yield the same background intensity level for normalization purposes. For each slide, a report with 4-channel images and preliminary prediction classifications of candidate cells was reviewed and signed off by an analyst (ASG).…”

Section: Hd-sca Workflow and Crc Assay Development And Validationmentioning

confidence: 99%

Untitled

2019

Oncotarget

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As cancer care is transitioning to personalized therapies with necessary complementary or companion biomarkers there is significant interest in determining to what extent non-invasive liquid biopsies reflect the gold standard solid biopsy. We have established an approach for measuring patient-specific circulating and solid cell concordance by introducing tumor touch preparations to the High-Definition Single Cell Analysis workflow for high-resolution cytomorphometric characterization of metastatic colorectal cancer (mCRC). Subgroups of cells based on size, shape and protein expression were identified in both liquid and solid biopsies, which overall displayed high inter-and intra-patient pleomorphism at the single-cell level of analysis. Concordance of liquid and solid biopsies was patient-dependent and between 0.1-0.9. Morphometric variables displayed particularly high correlation, suggesting that circulating cells do not represent distinct subpopulations from the solid tumor. This was further substantiated by significant decrease in concentration of circulating cells after mCRC resection. Combined with the association of circulating cells with tumor burden and necrosis of hepatic lesions, our overall findings demonstrate that liquid biopsy cells can be informative biomarkers in the mCRC setting. Patient-specific level of concordance can readily be measured to establish the utility of circulating cells as biomarkers and define biosignatures for liquid biopsy assays.

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Current Status of Circulating Tumor Cells in Head and Neck Squamous Cell Carcinoma: A Review

Aktar

Baghaie

Islam

et al. 2023

Otolaryngol.--head neck surg.

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Objective Circulating tumor cells (CTCs) are found in the blood of patients with cancer, including head and neck squamous cell carcinomas (HNSCCs). The aim is to review the most up‐to‐date status of CTCs for applications in patients with HNSCC. Data Sources English articles in PubMed. Review Methods All the studies on CTCs in HNSCCs in the literature were reviewed. Conclusions There is emerging information on the diagnostic and prognostic value of CTCs in HNSCCs. Evidence also highlights the advantages of various downstream analysis approaches over circulating tumor DNA (ctDNA), such as single‐CTC analysis, ex vivo, and in vivo expansion of CTCs. Multiple phenotypic surface markers (cytokeratins, EpCAM, vimentin, etc.), used for CTCs characterization using different immunoassays, could predict disease progression as well as patients' response to treatment efficacy. Immune checkpoint inhibitors' status in CTCs could also provide better insight into treatment. Clonal expansion of CTCs and single‐cell analysis of CTCs are the most emerging fields nowadays which may offer an understanding of the mechanism of tumor evolution as well as therapeutic efficacy. Although several clinical trials are ongoing, limitations still exist in the detection and characterization of CTCs. Due to the lack of a gold standard protocol, the sensitivity and specificity of CTC enumeration methods vary. Implications for Practice Prospective clinical trials are still needed before CTCs can be employed as diagnostic and prognostic markers in the clinical management of patients with HNSCC.

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Effect of Blood Collection Tube Type and Time to Processing on the Enumeration and High-Content Characterization of Circulating Tumor Cells Using the High-Definition Single-Cell Assay

Cited by 44 publications

References 25 publications

Liquid biopsies

Liquid biopsies

Untitled

Current Status of Circulating Tumor Cells in Head and Neck Squamous Cell Carcinoma: A Review

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