Circulating tumor cell

82:, presents several advantages over traditional tissue biopsies. They are non-invasive, can be used repeatedly, and provide more useful information on metastatic risk, disease progression, and treatment effectiveness. For example, analysis of blood samples from cancer patients has found a propensity for increased CTC detection as the disease progresses. Blood tests are easy and safe to perform and multiple samples can be taken over time. By contrast, analysis of solid tumors necessitates invasive procedures that might limit patient compliance. The ability to monitor the disease progression over time could facilitate appropriate modification to a patient's therapy, potentially improving their prognosis and quality of life. The important aspect of the ability to prognose the future progression of the disease is elimination (at least temporarily) of the need for a surgery when the repeated CTC counts are low and not increasing; the obvious benefits of avoiding the surgery include avoiding the risk related to the innate tumor-genicity of cancer surgeries. To this end, technologies with the requisite sensitivity and reproducibility to detect CTCs in patients with metastatic disease have recently been developed. On the other hand, CTCs are very rare, often present as only a few cells per milliliter of blood, which makes their detection challenging. In addition, they often express a variety of markers which vary from patient to patient, which makes it difficult to develop techniques with high 369:

process, Maintrac uses just two steps towards the identification. In contrast to many other methods, maintrac does not purify the cells or enrich them, but identifies them within the context of the other blood compounds. To obtain vital cells and to reduce stress of those cells, blood cells are prepared by only one centrifugation step and erythrocyte lysis. Like CellSearch, maintrac uses an EpCAM antibody. It is, however, not used for enrichment but rather as a fluorescent marker to identify those cells. Together with the nuclear staining with propidium iodide the maintrac method can distinguish between dead and living cells. Only vital, propidium excluding EpCAM positive cells are counted as potential tumor cells. Only living cells can grow into tumors, therefore dying EpCAM positive cells can do no harm. The suspension is analysed by fluorescence microscopy, which automatically counts the events. Simultaneous event galleries are recorded to verify whether the software found a true living cell and to differentiate between skin epithelial cells for example. Close validation of the method showed that additional antibodies of cytokeratins or CD45 did not have any advantage.

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blood cells and white blood cells to pass through, while larger rare cells, including circulating tumor cells and fetal cells get caught. Trapped cells can either be automatically stained with antibodies for identification or can be released out of the cassette for subsequent analysis. These released / harvested cells are alive and can be analyzed by downstream cellular and molecular techniques, as well as cultured. The filtration cassette captures a plethora of different cancer cell types. In May 2022, the Parsortix PC1 system was cleared by the FDA as a medical device for the capture and harvest of circulating tumor cells (CTCs) from metastatic breast cancer patient blood for subsequent analysis. In addition to the IVD application, the PC1 may be used with the MBC-01 Metastatic Breast Cancer Kit for use in research studies or Lab Developed Tests (LDTs) that have been created and validated in a clinical laboratory.

325:(epithelial markers). The sample is then scanned on an analyzer which takes images of the nuclear, cytokeratin, and CD45 stains. To be considered a CTC a cell must contain a nucleus, be positive for cytoplasmic expression of cytokeratin as well as negative for the expression of CD45 marker, and have a diameter larger than 5 μm. If the total number of tumor cells found to meet the criteria cited above is 5 or more, a blood sample is positive. In studies done on prostate, breast and colon cancer patients, median survival of metastatic patients with positive samples is about half the median survival of metastatic patients with negative samples. This system is characterized by a recovery capacity of 93% and a detection limit of one CTC per 7.5 mL of whole blood. For specific cancer types, alternative methods such as IsoFlux have shown greater 236:

total cells in circulating blood, 1–10 CTCs per mL of whole blood compared to a few million white blood cells and a billion red blood cells. Therefore, the major challenge for CTC researchers is the prevailing difficulty of CTC purification that allows the molecular characterization of CTCs. Several methods have been developed to isolate CTCs in the peripheral blood and essentially fall into two categories: biological methods and physical methods, as well as hybrid methods that combine both strategies. Techniques may also be classified based on whether they select CTCs for isolation (positive selection) or whether they exclude all blood cells (negative selection).

284:, as developed by GILUPI GmbH. An antibody coated metal wire is inserted into a peripheral vein and stays there for a defined period (30 min). During this time, CTCs from the blood can bind to the antibodies (currently anti-EpCAM). After the incubation time, the wire is removed, washed and the native CTCs, isolated from the blood of the patient, can be further analysed. Molecular genetics as well as immunofluorescent staining and several other methods are possible. Advantage of this method is the higher blood volume that can be analysed for CTCs (approx. 750 ml in 30 min compared to 7.5 ml of a drawn blood sample). 180:

cancerous cells, means that a key component of understanding CTCs biological properties require technologies and approaches capable of isolating 1 CTC per mL of blood, either by enrichment, or better yet with enrichment-free assays that identify all CTC subtypes in sufficiently high definition to satisfy diagnostic pathology image-quantity requirements in patients with a variety of cancer types. To date CTCs have been detected in several epithelial cancers (breast, prostate, lung, and colon) and clinical evidences indicate that patients with metastatic lesions are more likely to have CTCs isolated.

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next-generation sequencing. A hematopathology-trained algorithm incorporates numerous morphology measurements as well as expression from cytokeratin and CD45. The algorithm then proposes candidate CTCs that a trained reader confirms. Cells of interest are analyzed for relevant phenotypic and genotypic markers, with regional white blood cells included as negative controls. Epic's molecular assays measure protein expression and also interrogate genomic abnormalities in CTCs for more than 20 different cancer types.

20: 394:. ScreenCell is a filtration based device that allows sensitive and specific isolation of CTCs from human whole blood in a few minutes. Peripheral blood is drawn and processed within 4 hours with a ScreenCell isolation device to capture CTCs. The captured cells are ready for cell culture or for direct characterization using ViewRNA in situ hybridization assay. The Parsortix method separates CTCs based on their size and deformability. 503:(colloidal magnetic nanoparticles) and high gradient magnetic separators invented by Paul Liberti and motivated by theoretical calculations by Liberti and Leon Terstappen that indicated very small tumors shedding cells at less than 1.0% per day should result in detectable cells in blood. A variety of other technologies have been applied to CTC enumeration and identification since that time. 216: 151:

risk and poor prognosis. For example, one study involving prostate cancer showed an eight-fold longer mean survival rate for patients with only single CTCs versus those with CTC clusters, while other studies have shown similar correlations for colon cancer. In addition, enumerating CTC clusters can provide useful prognostic information for patients with already elevated CTC levels.

137:(programmed cell death). These may be used to monitor treatment response, as done experimentally by the Epic Sciences method, which identifies nuclear fragmentation or cytoplasmic blebbing associated with apoptosis. Measuring the ratio of traditional CTC to apoptotic CTCs—from baseline to therapy—provides clues to treatment efficacy in targeting and killing cancer cells. 373:

maintrac can be used to verify the success of a chemotherapy and to supervise the treatment during hormone or maintenance therapy Maintrac has been used experimentally to monitor cancer recurrence. Studies using Maintrac have shown that EpCAM positive cells can be found in the blood in patient without cancer. Inflammatory conditions like

264:. Other techniques under research include microfluidic separation and combination of immunomagnetic assay and microfluidic separation. As the development of microfabrication technology, microscale magnetic structures are implemented to provide better control of the magnetic field and assist the CTCs detection. Oncolytic viruses such as 63:) in distant organs, a mechanism that is responsible for the vast majority of cancer-related deaths. The detection and analysis of CTCs can assist early patient prognoses and determine appropriate tailored treatments. Currently, there is one FDA-approved method for CTC detection, CellSearch, which is used to diagnose 368:

diagnostic methods to identify rare cells in body fluids and their molecular characteristics. It is based on positive selection using EpCAM-specific antibodies. Maintrac uses an approach based on microscopic identification of circulating tumor cells. To prevent damage and loss of the cells during the

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tube with an added preservative. Upon arrival in the lab, 7.5mL of blood is centrifuged and placed in a preparation system. This system first enriches the tumor cells immunomagnetically by means of ferrofluid nanoparticles and a magnet. Subsequently, recovered cells are permeabilized and stained with

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CTC clusters are two or more individual CTCs bound together. The CTC cluster may contain traditional, small or CK- CTCs. These clusters have cancer-specific biomarkers that identify them as CTCs. Several studies have reported that the presence of these clusters is associated with increased metastatic

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of the circulating cells to tumor cells from different lesions led Ashworth to conclude that "One thing is certain, that if they came from an existing cancer structure, they must have passed through the greater part of the circulatory system to have arrived at the internal saphena vein of the sound

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CTCs are pivotal to understanding the biology of metastasis and promise potential as a biomarker to noninvasively evaluate tumor progression and response to treatment. However, isolation and characterization of CTCs represent a major technological challenge, since CTCs make up a minute number of the

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However, one study has reported that contrary to existing consensus, at least a discrete population of these clusters are non-malignant, and derive instead from the tumor endothelium. These circulating tumor-endothelial clusters also show epithelial-mesenchymal markers but do not mirror the genetics

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Lobodasch, Kurt; Fröhlich, Frank; Rengsberger, Matthias; Schubert, Rene; Dengler, Robert; Pachmann, Ulrich; Pachmann, Katharina (April 2007). "Quantification of circulating tumour cells for the monitoring of adjuvant therapy in breast cancer: An increase in cell number at completion of therapy is a

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Cima, I.; Kong, S. L.; Sengupta, D.; Tan, I. B.; Phyo, W. M.; Lee, D.; Hu, M.; Iliescu, C.; Alexander, I.; Goh, W. L.; Rahmani, M.; Suhaimi, N.-A. M.; Vo, J. H.; Tai, J. A.; Tan, J. H.; Chua, C.; Ten, R.; Lim, W. J.; Chew, M. H.; Hauser, C. A. E.; van Dam, R. M.; Lim, W.-Y.; Prabhakar, S.; Lim, B.;

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Morphological appearance is judged by human operators and is therefore subject to large inter operator variation. Several CTC enumeration methods exist which use morphological appearance to identify CTC, which may also apply different morphological criteria. A recent study in prostate cancer showed

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showed they could generate bone, lung, ovary and brain metastases in mice, partially reflecting the secondary lesions as found in the corresponding patients. Remarkably, one CTC line—isolated long before the appearance of brain metastasis in patient—was highly competent to generate brain metastasis

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This method involves technology to separate nucleated cells from red blood cells, which lack a nucleus. All nucleated cells, including normal white blood cells and CTCs, are exposed to fluorescent-tagged antibodies specific for cancer biomarkers. In addition, Epic's imaging system captures pictures

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Agerbæk, Mette Ø.; Bang-Christensen, Sara R.; Yang, Ming-Hsin; Clausen, Thomas M.; Pereira, Marina A.; Sharma, Shreya; Ditlev, Sisse B.; Nielsen, Morten A.; Choudhary, Swati; Gustavsson, Tobias; Sorensen, Poul H.; Meyer, Tim; Propper, David; Shamash, Jonathan; Theander, Thor G.; Aicher, Alexandra;

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Ferraldeschi, Roberta; McDaniel, Andrew; Krupa, Rachel; Louw, Jessica; Tucker, Eric; Bales, Natalee; Marrinucci, Dena; Riisnaes, Ruth; Mateo, Joaquin; Dittamore, Ryan; De Bono, Johann Sebastian; Tomlins, Scott A.; Attard, Gerhardt (February 2014). "CK- and small nuclear size circulating tumor cell

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Unlike other methods maintrac does not use the single cell count as a prognostic marker, rather Maintrac utilizes the dynamics of the cell count. Rising tumor cell numbers are an important factor that tumor activity is ongoing. Decreasing cell counts are a sign for a successful therapy. Therefore,

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This automated method uses size filtration to enrich larger and less compressible circulating tumor cells from other blood components. The Parsortix system can take in blood samples ranging from 1 mL to 40 mL. A disposable microfluidic cassette with a 6.5 micron gap allows the vast majority of red

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First evidence indicates that CTC markers applied in human medicine are conserved in other species. Five of the more common markers including CK19 are also useful to detect CTC in the blood of dogs with malignant mammary tumors. Newer approaches are able to identify more cells out 7.5 ml of blood,

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Sánchez-Lorencio, M.I.; Ramirez, P.; Saenz, L.; Martínez Sánchez, M.V.; De La Orden, V.; Mediero-Valeros, B.; Veganzones-De-Castro, S.; Baroja-Mazo, A.; Revilla Nuin, B.; Gonzalez, M.R.; Cascales-Campos, P.A.; Noguera-Velasco, J.A.; Minguela, A.; Díaz-Rubio, E.; Pons, J.A.; Parrilla, P. (November

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Pachmann K.; Camara O.; Kavallaris A.; Krauspe S.; Malarski N.; Gajda M.; Kroll T.; Jorke C.; Hammer U.; Altendorf-Hofmann A.; et al. (2008). "Monitoring the Response of Circulating Epithelial Tumor Cells to Adjuvant Chemotherapy in Breast Cancer Allows Detection of Patients at Risk of Early

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CTCs were observed for the first time in 1869 in the blood of a man with metastatic cancer by Thomas Ashworth, who postulated that "cells identical with those of the cancer itself being seen in the blood may tend to throw some light upon the mode of origin of multiple tumours existing in the same

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Small CTCs are cytokeratin-positive and CD45-negative, but with sizes and shapes similar to white blood cells. Importantly, small CTCs have cancer-specific biomarkers that identify them as CTCs. Small CTCs have been implicated in progressive disease and differentiation into small cell carcinomas,

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Ozkumur E, Shah AM, Ciciliano JC, Emmink BL, Miyamoto DT, Brachtel E, Yu M, Chen PI, Morgan B, Trautwein J, Kimura A, Sengupta S, Stott SL, Karabacak NM, Barber TA, Walsh JR, Smith K, Spuhler PS, Sullivan JP, Lee RJ, Ting DT, Luo X, Shaw AT, Bardia A, Sequist LV, Louis DN, Maheswaran S, Kapur R,

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Divella R, Daniele A, Abbate I, Bellizzi A, Savino E, Simone G, Giannone G, Giuliani F, Fazio V, Gadaleta-Caldarola G, Gadaleta C, Lolli I, Sabbà C, Mazzocca A (2014). "The presence of clustered circulating tumor cells (CTCs) and circulating cytokines define an aggressive phenotype in metastatic

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positive, but much less effective in patients who are Her2 negative. Once the primary tumor is removed, biopsy of the current state of the cancer through traditional tissue typing is not possible anymore. Often tissue sections of the primary tumor, removed years prior, are used to do the typing.

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Hybrid methods combine physical separation (by gradients, magnetic fields, etc.) with antibody-mediated cell retrieval. An example of this is a sensitive double gradient centrifugation and magnetic cell sorting detection and enumeration method which has been used to detect circulating epithelial

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Circulating tumor cells are found in frequencies on the order of 1-10 CTC per mL of whole blood in patients with metastatic disease. For comparison, a mL of blood contains a few million white blood cells and a billion red blood cells. This low frequency, associated to difficulty of identifying

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Marrinucci, Dena; Bethel, Kelly; Kolatkar, Anand; Luttgen, Madelyn; Malchiodi, Michael; Baehring, Franziska; Voigt, Katharina; Lazar, Daniel; Nieva, Jorge; Bazhenova, Lyudmilda; Ko, Andrew; Korn, W. Michael; Schram, Ethan; Coward, Michael; Yang, Xing; Metzner, Thomas; Lamy, Rachelle; Honnatti,

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Modern cancer research has demonstrated that CTCs derive from clones in the primary tumor, validating Ashworth's remarks. The significant efforts put into understanding the CTCs biological properties have demonstrated the critical role circulating tumor cells play in the metastatic spread of

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Previously it was assumed that CTC clusters could not pass through narrow vessels, such as capillaries, due to their overall size. However, it has been shown that CTC clusters can "unwind" through "selective cleavage of intercellular adhesions" to traverse these constrictions single-file, then

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Hodgkinson, Cassandra L; Morrow, Christopher J; Li, Yaoyong; Metcalf, Robert L; Rothwell, Dominic G; Trapani, Francesca; Polanski, Radoslaw; Burt, Deborah J; Simpson, Kathryn L; Morris, Karen; Pepper, Stuart D; Nonaka, Daisuke; Greystoke, Alastair; Kelly, Paul; Bola, Becky; Krebs, Matthew G;

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Kaplan Meier Analysis of overall survival before starting a new line of therapy for patients with metastatic breast, colorectal and prostate cancer. Patients were divided into those with Favorable and Unfavorable CTC (Unfavorable: >5 CTC/7.5mL for breast and prostate, >3 CTC/7.5mL for

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and other staining techniques to look for abnormalities such as duplications, deletions, and rearrangements. The imaging and analysis technology also allows for the coordinates of every cell on a slide to be known so that a single cell can be retrieved from the slide for analysis using

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of all the cells on the slide (approximately 3 million), records the precise coordinates of each cell, and analyzes each cell for 90 different parameters, including the fluorescence intensity of the four fluorescent markers and 86 different morphological parameters. Epic can also use

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Nagrath, Sunitha; Sequist, Lecia V.; Maheswaran, Shyamala; Bell, Daphne W.; Irimia, Daniel; Ulkus, Lindsey; Smith, Matthew R.; Kwak, Eunice L.; Digumarthy, Subba; Muzikansky, Alona; Ryan, Paula; Balis, Ulysses J.; Tompkins, Ronald G.; Haber, Daniel A.; Toner, Mehmet (December 2007).

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or EMT). These populations of CTCs may be the most resistant and most prone to metastasis. They are also more difficult to isolate because they express neither cytokeratins nor CD45. Otherwise, their morphology, gene expression and genomics are similar to those of other cancer

800: 175:

The detection of CTCs may have important prognostic and therapeutic implications but because their numbers can be very small, these cells are not easily detected. It is estimated that among the cells that have detached from the primary tumor, only 0.01% can form metastases.

511:. Furthermore, highly sensitive, single-cell analysis demonstrated a high level of heterogeneity seen at the single cell level for both protein expression and protein localization and the CTCs reflected both the primary biopsy and the changes seen in the metastatic sites. 4072:

Antonello, Jenny; Ayub, Mahmood; Faulkner, Suzanne; Priest, Lynsey; Carter, Louise; Tate, Catriona; Miller, Crispin J; Blackhall, Fiona; Brady, Ged; Dive, Caroline (1 June 2014). "Tumorigenicity and genetic profiling of circulating tumor cells in small-cell lung cancer".

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Fehm T, Sagalowsky A, Clifford E, Beitsch P, Saboorian H, Euhus D, Meng S, Morrison L, Tucker T, Lane N, Ghadimi BM, Heselmeyer-Haddad K, Ried T, Rao C, Uhr J (Jul 2002). "Cytogenetic evidence that circulating epithelial cells in patients with carcinoma are malignant".

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Attard G, Swennenhuis JF, Olmos D, Reid AH, Vickers E, A'Hern R, Levink R, Coumans F, Moreira J, Riisnaes R, Oommen NB, Hawche G, Jameson C, Thompson E, Sipkema R, Carden CP, Parker C, Dearnaley D, Kaye SB, Cooper CS, Molina A, Cox ME, Terstappen LW, de Bono JS (2009).

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that many different morphological definitions of circulating tumor cells have similar prognostic value, even though the absolute number of cells found in patients and normal donors varied by more than a decade between different morphological definitions.

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Yu M, Ting DT, Stott SL, Wittner BS, Ozsolak F, Paul S, Ciciliano JC, Smas ME, Winokur D, Gilman AJ, Ulman MJ, Xega K, Contino G, Alagesan B, Brannigan BW, Milos PM, Ryan DP, Sequist LV, Bardeesy N, Ramaswamy S, Toner M, Maheswaran S, Haber DA (2012).

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The importance of CTCs in modern cancer research began in the mid 1990s with the demonstration that CTCs exist early on in the course of the disease. Those results were made possible by exquisitely sensitive magnetic separation technology employing

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Meng S, Tripathy D, Shete S, Ashfaq R, Haley B, Perkins S, Beitsch P, Khan A, Euhus D, Osborne C, Frenkel E, Hoover S, Leitch M, Clifford E, Vitetta E, Morrison L, Herlyn D, Terstappen LW, Fleming T, Fehm T, Tucker T, Lane N, Wang J, Uhr J (2004).

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analogues and conjugated with antibodies against EpCAM for the capture of CTCs. Isolation is coupled to an analyzer to take images of isolated cells upon their staining with specific fluorescent antibody conjugates. Blood is sampled in an

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Luecke, Klaus, et al. "The GILUPI CellCollector as an in vivo tool for circulating tumor cell enumeration and molecular characterization in lung cancer patients." ASCO Annual Meeting Proceedings. Vol. 33. No. 15_suppl. 2015.

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in mice. This was the first predictive case for brain metastasis and a proof of concept that intrinsic molecular features of metastatic precursors amongst CTCs could provide novel insights into the mechanisms of metastasis.

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also show increased levels of EpCAM-positive cells. Patients with severe skin burns can also carry EpCAM positive cells in the blood. Therefore, the use of EpCAM-positive cells as a tool for early diagnosis is not optimal.

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Ye Z, Mu Z, Wang C, Palazzo JP, Biederman L, Li B, Jaslow R, Avery T, Austin L, Yang H, Cristofanilli M (2016). "Prognostic values of circulating tumor cell (CTC) enumeration and their clusters in advanced breast cancer".

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Polascik TJ, Wang ZP, Shue M, Di S, Gurganus RT, Hortopan SC, Ts'o PO, Partin AW (1999). "Influence of sextant prostate needle biopsy or surgery on the detection and harvest of intact circulating prostate cancer cells".

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Wang, Huiqiang; Chen, Nanhai G.; Minev, Boris R.; Zimmermann, Martina; Aguilar, Richard J.; Zhang, Qian; Sturm, Julia B.; Fend, Falko; Yu, Yong A.; Cappello, Joseph; Lauer, Ulrich M.; Szalay, Aladar A. (September 2013).

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Aceto N, Bardia A, Miyamoto DT, Donaldson MC, Wittner BS, Spencer JA, Yu M, Pely A, Engstrom A, Zhu H, Brannigan BW, Kapur R, Stott SL, Shioda T, Ramaswamy S, Ting DT, Lin CP, Toner M, Haber DA, Maheswaran S (2014).

3903:

Tkaczuk KH, Goloubeva O, Tait NS, Feldman F, Tan M, Lum ZP, Lesko SA, Van Echo DA, Ts'o PO (2008). "The significance of circulating epithelial cells in Breast Cancer patients by a novel negative selection method".

187:); however this approach is biased by the need for a sufficient expression of the selected protein on the cell surface, event necessary for the enrichment step. Moreover, since EpCAM and other proteins (e.g. 98:(cancers of epithelial origin, which are the most prevalent) can be classified according to the expression of epithelial markers, as well as their size and whether they are apoptotic. In general, CTCs are 228:(FDA) cleared methodology for enumeration of CTC in whole blood is the CellSearch system. Extensive clinical testing done using this method shows that presence of CTCs is a strong prognostic factor for 612:

Rack B, Schindlbeck C, Jückstock J, Andergassen U, Hepp P, Zwingers T, Friedl T, Lorenz R, Tesch H, Fasching P, Fehm T, Schneeweiss A, Lichtenegger W, Beckmann M, Friese K, Pantel K, Janni W (2014).

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Klotz, Remi; Thomas, Amal; Teng, Teng; Han, Sung Min; Iriondo, Oihana; Li, Lin; Restrepo-Vassalli, Sara; Wang, Alan; Izadian, Negeen; MacKay, Matthew; Moon, Byoung-San (2019-01-01).

3725:"Increase in number of circulating disseminated epithelial cells after surgery for non-small cell lung cancer monitored by MAINTRAC is a predictor for relapse: A preliminary report" 3264: 3233:"50P * Enumeration and Molecular Characterization of Circulating Tumor Cells in Lung Cancer Patients Using the Gilupi Cellcollector , an Effective in Vivo Device for Capturing CTCS" 2005:

da Costa A, Oliveira JT, Gärtner F, Kohn B, Gruber AD, Klopfleisch R (2011). "Potential markers for detection of circulating canine mammary tumor cells in the peripheral blood".

4635: 3576:"Assessing the efficacy of targeted therapy using circulating epithelial tumor cells (CETC): the example of SERM therapy monitoring as a unique tool to individualize therapy" 1870:

Tanaka F, Yoneda K, Kondo N, Hashimoto M, Takuwa T, Matsumoto S, Okumura Y, Rahman S, Tsubota N, Tsujimura T, Kuribayashi K, Fukuoka K, Nakano T, Hasegawa S (2009).

423:. The technique, called CTC-iChip, first removes cells too small to be CTCs, such as red blood cells, and then uses magnetic particles to remove white blood cells. 2681:

Peng, Chen; Yu-yen, Huang; Hoshino, Kazunori; Xiaojing, Zhang (2014). "Multiscale immunomagnetic enrichment of circulating tumor cells: from tubes to microchips".

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Cohen SJ, Punt CJ, Iannotti N, Saidman BH, Sabbath KD, Gabrail NY, Picus J, Morse M, Mitchell E, Miller MC, Doyle GV, Tissing H, Terstappen LW, Meropol NJ (2008).

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Huang, Yu-yen; Hoshino, Kazunori; Chen, Peng; Wu, Chun-hsien; Lane, Nancy; Huebschman, Michael; Liu, Huaying; Sokolov, Konstantin; Uhr, Jonathan W. (2012-10-30).

3164:"A novel method for the in vivo isolation of circulating tumor cells from peripheral blood of cancer patients using a functionalized and structured medical wire" 2903:

Huang, Yu-Yen; Chen, Peng; Wu, Chun-Hsien; Hoshino, Kazunori; Sokolov, Konstantin; Lane, Nancy; Liu, Huaying; Huebschman, Michael; Frenkel, Eugene (2015-11-05).

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Bethel, Kelly; Luttgen2, Madelyn; Damani, Samir; Kolatkar2, Anand; Lamy, Rachelle; Sabouri-Ghomi, Mohsen; Topol, Sarah; Topol2, Eric; Kuhn, Peter (9 Jan 2014).

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Nieva, J; Wendel, M; Luttgen, MS; Marrinucci, D; Bazhenova, L; Kolatkar, A; Santala, R; Whittenberger, B; Burke, J; Torrey, M; Bethel, K; Kuhn, P (Feb 2012).

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Hayes DF, Walker TM, Singh B, et al. (2002). "Monitoring Expression of HER-2 on Circulating Epithelial Cells in Patients with advanced Breast Cancer".

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Au S, Storey B, Moore J, Tang Q, Chen Y, Javaid S, Sarioglu A, Sullivan R, Madden M, O'Keefe R, Haber D, Maheswaran S, Langenau D, Stott S, Toner M (2016).

801:"Relationship of circulating tumor cells to tumor response, progression-free survival, and overall survival in patients with metastatic colorectal cancer" 3215: 3535:"The relevance of circulating epithelial tumor cells (CETC) for therapy monitoring during neoadjuvant (primary systemic) chemotherapy in breast cancer" 1107:

Esmaeilsabzali H, Beischlag TV, Cox ME, Parameswaran AM, Park EJ (2013). "Detection and isolation of circulating tumor cells: principles and methods".

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Further characterization of CTC may help determining the current tumor phenotype. FISH assays have been performed on CTC as well as determination of

3625:"Efficacy control of therapy using circulating epithelial tumor cells (CETC) as "Liquid Biopsy": trastuzumab in HER2/neu-positive breast carcinoma" 1147:"High-definition imaging of circulating tumor cells and associated cellular events in non-small cell lung cancer patients: a longitudinal analysis" 419:

patients. Similarly, researchers at Massachusetts General Hospital have developed a negative selection method which employs inertial focusing on a

3381: 2486:"Detection of Circulating Tumor Cells in Peripheral Blood of Patients with Metastatic Breast Cancer: A Validation Study of the CellSearch System" 896:

Sleijfer S, Gratama JW, Sieuwerts AM, et al. (2007). "Circulating tumour cell detection on its way to routine diagnostic implementation?".

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Cytokeratin-negative CTCs are characterised by the lack of EpCAM or cytokeratins, which may indicate an undifferentiated phenotype (circulating

3440:"Quantification of the response of circulating epithelial cells to neodadjuvant treatment for breast cancer: a new tool for therapy monitoring" 1721:"Significance of Circulating Tumor Cells Detected by the CellSearch System in Patients with Metastatic Breast Colorectal and Prostate Cancer" 415:

has been employed to detect circulating prostate cancer cells and another technique known as RosetteStep has been used to isolate CTCs from

2445:"Tumor cells circulate in the peripheral blood of all major carcinomas but not in healthy subjects or patients with non-malignant diseases" 3271: 3289:

2015). "Comparison of Two Types of Liquid Biopsies in Patients With Hepatocellular Carcinoma Awaiting Orthotopic Liver Transplantation".

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CTCs are usually (in 2011) captured from the vasculature by using specific antibodies able to recognize specific tumoral marker (usually

467:

The organ tropism of patient-derived CTC has been investigated in a mouse model. CTCs isolated from breast cancer patients and expanded

4344:"Pharmacokinetics and Pharmacodynamics of the IGF-IR Inhibitor Figitumumab (CP-751,871) in Combination with Paclitaxel and Carboplatin" 3395:

Pachmann, Katharina (5 April 2015). "Current and potential use of MAINTRAC method for cancer diagnosis and prediction of metastasis".

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Hoshino, Kazunori; Huang, Yu-Yen; Lane, Nancy; Huebschman, Michael; Uhr, Jonathan W.; Frenkel, Eugene P.; Zhang, Xiaojing (Oct 2011).

757:"Characterization of ERG, AR and PTEN gene status in circulating tumor cells from patients with castration-resistant prostate cancer" 260:

have been used. The most common technique is magnetic nanoparticle-based separation (immunomagnetic assay) as used in CellSearch or

4024:

Ali A, Furusato B, Ts'o PO, Lum ZP, Elsamanoudi S, Mohamed A, Srivastava S, Moul JW, Brassell SA, Sesterhenn IA, McLeod DG (2010).

2283:"Veridex LLC. CellSearch circulating tumor cell kit premarket notification—expanded indications for use—metastatic prostate cancer" 1648:

Ghossein RA, Bhattacharya S, Rosai J (1999). "Molecular detection of micrometastases and circulating tumor cells in solid tumors".

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cancer cells in breast cancer patients by negative selection. The principle of negative selection is based on the retrieval of all

3825:

Desitter I.; et al. (2011). "A New Device for Rapid Isolation by Size and Characterization of Rare Circulating Tumor Cells".

2257: 2222:

Paterlini-Brechot P, Benali NL.; Benali (2007). "Circulating tumor cells (CTC) detection: Clinical impact and future directions".

2040:

da Costa, A (2013). "Multiple RT-PCR markers for the detection of circulating tumour cells of metastatic canine mammary tumours".

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reverse the process once clear. This behavior could be a factor in why CTC clusters have such a significant metastatic potential.

3965: 3948: 3776:"Seeding of Epithelial Cells into Circulation During Surgery for Breast Cancer: The Fate of Malignant and Benign Mobilized Cells" 2404:"Circulating Tumor Cells (CTC) predict survival benefit from treatment in metastatic castration resistant prostate cancer (CRPC)" 114:, which demonstrate epithelial origin; the absence of CD45, indicating the cell is not of hematopoietic origin; and their larger 2175:"Carcinocythemia: A rare entity becoming more common? A 3-year, single institution series of seven cases and literature review" 4540:

Ashworth, T. R (1869). "A case of cancer in which cells similar to those in the tumours were seen in the blood after death".

441: 192: 704:"Circulating tumor cells from well-differentiated lung adenocarcinoma retain cytomorphologic features of primary tumor type" 3674:"Epithelial Cell Dissemination and Readhesion: Analysis of Factors Contributing to Metastasis Formation in Breast Cancer" 3533:

Camara O.; Rengsberger M.; Egbe A.; Koch A.; Gajda M.; Hammer U.; Jorke C.; Rabenstein C.; Untch M.; Pachmann K. (2007).

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3774:

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like IsofFux or Maintrac. In very rare cases, CTCs are present in large enough quantities to be visible on routine

1060:"Detection of circulating tumor cells in prostate cancer patients: methodological pitfalls and clinical relevance" 3326:"Fluid phase biopsy for detection and characterization of circulating endothelial cells in myocardial infarction" 261: 4262:"Multigene Reverse Transcription-PCR Profiling of Circulating Tumor Cells in Hormone-Refractory Prostate Cancer" 23:

An illustration depicting primary tumor (in the form of tumor microenvironment) and the circulating tumor cells.

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191:) are not expressed in some tumors and can be down regulated during the epithelial to mesenchymal transition ( 3212: 1316:

Meghana; Yoshioka, Craig; Kunken, Joshua; Petrova, Yelena; Sok, Devin; Nelson, David; Kuhn, Peter (Feb 2012).

4771: 2840:"Microscale Magnetic Field Modulation for Enhanced Capture and Distribution of Rare Circulating Tumor Cells" 2484:

Riethdorf; Fritsche, H; Müller, V; Rau, T; Schindlbeck, C; Rack, B; Janni, W; Coith, C; et al. (2007).

2077:"Mutational Analysis of Circulating Tumor Cells Using a Novel Microfluidic Collection Device and qPCR Assay" 4628:"The tools that fueled the new world of Circulating Tumor Cells by Paul A. Liberti – BioMagnetic Solutions" 2290: 225: 3088:"The VAR2CSA malaria protein efficiently retrieves circulating tumor cells in an EpCAM-independent manner" 2783:"Computational Analysis of Microfluidic Immunomagnetic Rare Cell Separation from a Particulate Blood Flow" 4559:

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4501:"All circulating EpCAM1CK1CD452 objects predict overall survival in castration-resistant prostate cancer" 2970:"Inkjet-Print Micromagnet Array on Glass Slides for Immunomagnetic Enrichment of Circulating Tumor Cells" 326: 83: 4118:"Inertial focusing for tumor antigen-dependent and -independent sorting of rare circulating tumor cells" 663:"Circulating tumor cells: a review of present methods and the need to identify heterogeneous phenotypes" 292:

CellSearch is the only FDA-approved platform for CTC isolation. This method is based on the use of iron

224:

To date, a variety of research methods have been developed to isolate and enumerate CTCs. The only U.S.

3028:"Optical Detection and Virotherapy of Live Metastatic Tumor Cells in Body Fluids with Vaccinia Strains" 2726:"Immunomagnetic nanoscreening of circulating tumor cells with a motion controlled microfluidic system" 2173:

Ronen, Shira; Kroft, Steven H.; Olteanu, Horatiu; Hosking, Paul R.; Harrington, Alexandra M. (2019).

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2075:

Harb, W.; Fan, A.; Tran, T.; Danila, D.C.; Keys, D.; Schwartz, M.; and Ionescu-Zanetti, C. (2013).

431:

Some drugs are particularly effective against cancers which fit certain requirements. For example,

102:-resistant, which means that they can survive in the bloodstream without attaching to a substrate. 3574:

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841:"RNA sequencing of pancreatic circulating tumour cells implicates WNT signalling in metastasis" 614:"Circulating Tumor Cells Predict Survival in Early Average-to-High Risk Breast Cancer Patients" 416: 3375: 245: 2905:"Screening and Molecular Analysis of Single Circulating Tumor Cells Using Micromagnet Array" 244:

Biological methods isolate cells based on highly specific antigen binding, most commonly by

4572: 4179: 3337: 3099: 3039: 2916: 2851: 2588: 1602: 1525: 1329: 1264: 1158: 852: 491: 2968:

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8: 420: 281: 280:

on the surface of CTCs. CTCs may also be retrieved directly from the blood by a modified

277: 269: 4576: 4393:"Circulating tumor cells exhibit metastatic tropism and reveal brain metastasis drivers" 4183: 3349: 3341: 3103: 3043: 2920: 2855: 2592: 1606: 1404:"Circulating tumor cell clusters are oligoclonal precursors of breast cancer metastasis" 1333: 1268: 1162: 856: 4710: 4481: 4427: 4392: 4368: 4343: 4142: 4117: 4097: 4053: 3929: 3880: 3853: 3802: 3775: 3751: 3724: 3700: 3673: 3649: 3624: 3600: 3575: 3466: 3439: 3438:

Pachmann K.; Camara O.; Kavallaris A.; Schneider U.; Schünemann S.; Höffken K. (2005).

3420: 3358: 3325: 3302: 3188: 3163: 3120: 3087: 3062: 3027: 3002: 2969: 2945: 2904: 2880: 2839: 2815: 2782: 2758: 2725: 2706: 2658: 2633: 2609: 2577:"Isolation of rare circulating tumour cells in cancer patients by microchip technology" 2576: 2551: 2526: 2343: 2314:"Circulating Tumor Cells, Disease Progression and Survival in Metastatic Breast Cancer" 2155: 2101: 2076: 1982: 1955: 1936: 1847: 1822: 1798: 1771: 1747: 1720: 1693: 1676: 1625: 1590: 1571: 1509: 1477: 1428: 1403: 1350: 1341: 1317: 1228: 1203: 1179: 1170: 1146: 1084: 1059: 1040: 997: 873: 840: 728: 703: 679: 662: 638: 613: 594: 461: 374: 268:

viruses are developed to detect and identify CTCs. Alternative methods exist which use

48: 4202: 4167: 4776: 4702: 4667: 4608: 4603: 4590: 4560: 4522: 4473: 4432: 4414: 4373: 4324: 4283: 4242: 4207: 4147: 4101: 4089: 4057: 4045: 4041: 4006: 4002: 3970: 3921: 3885: 3834: 3807: 3756: 3705: 3654: 3605: 3556: 3515: 3507: 3471: 3412: 3363: 3306: 3193: 3125: 3067: 3007: 2989: 2950: 2932: 2885: 2867: 2820: 2802: 2763: 2745: 2710: 2698: 2663: 2614: 2556: 2507: 2466: 2425: 2384: 2335: 2239: 2204: 2196: 2147: 2106: 2057: 2022: 1987: 1928: 1893: 1852: 1803: 1752: 1698: 1657: 1630: 1563: 1555: 1469: 1433: 1355: 1292: 1287: 1252: 1233: 1184: 1124: 1120: 1089: 1032: 989: 954: 913: 878: 820: 778: 733: 684: 643: 586: 545: 457: 122: 68: 4728: 3933: 3424: 2159: 1940: 1575: 1481: 1387: 4714: 4694: 4598: 4580: 4512: 4485: 4463: 4422: 4404: 4363: 4355: 4314: 4273: 4234: 4197: 4187: 4137: 4129: 4081: 4037: 3998: 3960: 3913: 3875: 3865: 3797: 3787: 3746: 3736: 3695: 3685: 3644: 3636: 3595: 3587: 3546: 3499: 3461: 3451: 3404: 3353: 3345: 3298: 3244: 3183: 3175: 3115: 3107: 3057: 3047: 2997: 2981: 2940: 2924: 2875: 2859: 2810: 2794: 2753: 2737: 2690: 2653: 2645: 2604: 2596: 2546: 2538: 2497: 2456: 2415: 2374: 2347: 2325: 2261: 2231: 2186: 2139: 2096: 2088: 2049: 2014: 1977: 1967: 1920: 1883: 1842: 1834: 1793: 1783: 1742: 1732: 1688: 1620: 1610: 1545: 1537: 1505: 1461: 1423: 1415: 1383: 1345: 1337: 1282: 1272: 1223: 1215: 1174: 1166: 1116: 1079: 1071: 1044: 1024: 981: 944: 905: 868: 860: 812: 768: 723: 715: 674: 633: 625: 598: 576: 537: 407:

by using a panel of antibodies as well as traditional gradient centrifugation with

106:

Traditional CTCs are characterised by an intact, viable nucleus; the expression of

40: 4319: 4302: 3408: 2502: 2485: 2461: 2444: 2420: 2403: 2379: 2362: 1888: 1871: 1838: 1318:"Fluid Biopsy in Patients with Metastatic Prostate, Pancreatic and Breast Cancers" 1001: 949: 932: 773: 756: 464:. Single cell level qPCR can also be performed with the CTCs isolated from blood. 4409: 4359: 4278: 4261: 4133: 3503: 3219: 3052: 2235: 1541: 933:"Is There a Role for Circulating Tumor Cells in the Management of Breast Cancer?" 541: 361: 204: 72: 3671: 3249: 3232: 3111: 2053: 2018: 1419: 1374:(CTCs) phenotypes in metastatic castration-resistant prostate cancer (mCRPC)". 909: 581: 564: 248:

for positive selection. Antibodies against tumor specific biomarkers including

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Physical methods are often filter-based, enabling the capture of CTCs by

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or carcinoma cell leukemia and is associated with a poor prognosis.

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Number of various types of blood cells in whole blood versus CTCs.

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a nuclear stain, a fluorescent antibody conjugate against

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Apoptotic CTCs are traditional CTCs that are undergoing

4260:

O'Hara SM, Moreno JG, Zweitzig DR, et al. (2004).

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Budd G, Cristofanilli M, Ellis M, et al. (2006).

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Cell from a primary tumor carried by blood circulation

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Oncol 3112:10.1038/s41467-018-05793-2 2054:10.1016/j.tvjl.2012.08.021 2019:10.1016/j.tvjl.2010.09.027 1504:(4 Supplement): P2–08–09. 1420:10.1016/j.cell.2014.07.013 910:10.1016/j.ejca.2007.09.016 582:10.1016/j.cell.2006.11.001 484: 78:The detection of CTCs, or 4632:biomargneticsolutions.com 3918:10.1007/s10549-007-9771-9 3641:10.1007/s00432-011-1000-6 3592:10.1007/s00432-010-0942-4 2986:10.1007/s10439-015-1427-z 2742:10.1007/s10544-012-9718-8 1925:10.1007/s11864-010-0115-3 1466:10.1007/s10552-014-0457-4 1076:10.2119/molmed.2008.00116 1029:10.1038/nrclinonc.2009.23 986:10.1038/nrclinonc.2009.44 94:CTCs that originate from 4660:Clinical Cancer Research 3906:Breast Cancer Res. Treat 3250:10.1093/annonc/mdv045.14 2144:10.1200/JCO.2007.13.6523 1827:Clinical Cancer Research 817:10.1200/JCO.2007.15.8923 390:rather than by specific 89: 4752:www.abstractsonline.com 4733:meetinglibrary.asco.org 4193:10.1073/pnas.0402993101 2730:Biomedical Microdevices 1616:10.1073/pnas.1524448113 310:(leukocyte marker) and 4586:10.1073/pnas.95.8.4589 3793:10.1186/1477-7819-4-67 3742:10.1186/1477-7819-3-18 1789:10.1186/1741-7015-9-43 1278:10.1073/pnas.95.8.4589 417:small-cell lung cancer 221: 172: 155:of the primary tumor. 29:circulating tumor cell 24: 4687:Nature Reviews Cancer 4638:on September 26, 2017 4518:10.1093/annonc/mdq030 4239:10.3892/ijo.21.5.1111 3552:10.1093/annonc/mdm206 3180:10.3892/ijo.2012.1557 3092:Nature Communications 2543:10.1083/jcb.201010021 1454:Cancer Causes Control 1220:10.3892/ijo.2016.3747 362:diagnostic blood test 246:monoclonal antibodies 218: 170: 22: 4772:Anatomical pathology 4469:10.1002/cyto.a.20369 3871:10.1002/cyto.a.23571 3729:World J. Surg. Oncol 2787:Analytical Chemistry 2331:10.1056/NEJMoa040766 1452:colorectal cancer". 427:CTC characterization 342:Epic Sciences method 4577:1998PNAS...95.4589R 4184:2004PNAS..101.9393M 3827:Anticancer Research 3691:10.5402/2012/601810 3342:2014PhBio..11a6002B 3104:2018NatCo...9.3279A 3044:2013PLoSO...871105W 2921:2015NatSR...516047H 2856:2015NatSR...5E8745C 2601:10.1038/nature06385 2593:2007Natur.450.1235N 2587:(7173): 1235–1239. 1973:10.1155/2011/252361 1960:Journal of Oncology 1738:10.1155/2010/617421 1607:2016PNAS..113.4947A 1334:2012PhBio...9a6003M 1269:1998PNAS...95.4589R 1163:2012PhBio...9a6004N 865:10.1038/nature11217 857:2012Natur.487..510Y 630:10.1093/jnci/dju066 421:microfluidic device 282:Seldinger technique 278:chondroitin sulfate 270:engineered proteins 4505:Annals of Oncology 3997:(3 Pt 1): 749–52. 3237:Annals of Oncology 3218:2016-03-10 at the 2909:Scientific Reports 2844:Scientific Reports 2695:10.1039/C3LC51107C 2650:10.1039/c1lc20270g 2192:10.1111/ijlh.12924 2042:Veterinary Journal 974:Nat Rev Clin Oncol 720:10.5858/133.9.1468 462:immunofluorescence 240:Biological methods 222: 173: 25: 4403:(1): CD–19–0384. 4354:(11): 1397–1403. 3864:(12): 1234–1239. 3444:Breast Cancer Res 2929:10.1038/srep16047 2864:10.1038/srep08745 2799:10.1021/ac2032386 2793:(10): 4292–4299. 2644:(20): 3449–3457. 2455:(20): 6897–6904. 2093:10.1593/tlo.13367 288:CellSearch method 211:Detection methods 123:cancer stem cells 49:blood circulation 4784: 4756: 4755: 4743: 4737: 4736: 4725: 4719: 4718: 4682: 4676: 4675: 4654: 4648: 4647: 4645: 4643: 4634:. Archived from 4623: 4617: 4616: 4606: 4588: 4571:(8): 4589–4594. 4556: 4550: 4549: 4537: 4531: 4530: 4520: 4496: 4490: 4489: 4471: 4456:Cytometry Part A 4447: 4441: 4440: 4430: 4412: 4397:Cancer Discovery 4388: 4382: 4381: 4371: 4339: 4333: 4332: 4322: 4298: 4292: 4291: 4281: 4257: 4251: 4250: 4222: 4216: 4215: 4205: 4195: 4162: 4156: 4155: 4145: 4112: 4106: 4105: 4068: 4062: 4061: 4021: 4015: 4014: 3985: 3979: 3978: 3968: 3944: 3938: 3937: 3900: 3894: 3893: 3883: 3873: 3858:Cytometry Part A 3849: 3843: 3842: 3822: 3816: 3815: 3805: 3795: 3771: 3765: 3764: 3754: 3744: 3720: 3714: 3713: 3703: 3693: 3669: 3663: 3662: 3652: 3635:(9): 1317–1327. 3620: 3614: 3613: 3603: 3571: 3565: 3564: 3554: 3545:(9): 1484–1492. 3530: 3524: 3523: 3486: 3480: 3479: 3469: 3459: 3435: 3429: 3428: 3392: 3386: 3385: 3379: 3371: 3361: 3330:Physical Biology 3321: 3315: 3314: 3297:(9): 2639–2642. 3285: 3279: 3278: 3276: 3270:. Archived from 3269: 3261: 3255: 3254: 3252: 3228: 3222: 3208: 3202: 3201: 3191: 3174:(4): 1241–1250. 3159: 3153: 3152: 3140: 3134: 3133: 3123: 3082: 3076: 3075: 3065: 3055: 3022: 3016: 3015: 3005: 2980:(5): 1710–1720. 2965: 2959: 2958: 2948: 2900: 2894: 2893: 2883: 2835: 2829: 2828: 2818: 2778: 2772: 2771: 2761: 2721: 2715: 2714: 2678: 2672: 2671: 2661: 2629: 2623: 2622: 2612: 2571: 2565: 2564: 2554: 2522: 2516: 2515: 2505: 2481: 2475: 2474: 2464: 2440: 2434: 2433: 2423: 2399: 2393: 2392: 2382: 2358: 2352: 2351: 2333: 2309: 2303: 2302: 2300: 2299: 2287: 2279: 2273: 2272: 2270: 2269: 2254: 2248: 2247: 2219: 2213: 2212: 2194: 2170: 2164: 2163: 2138:(8): 1208–1215. 2126: 2115: 2114: 2104: 2072: 2066: 2065: 2037: 2031: 2030: 2002: 1996: 1995: 1985: 1975: 1951: 1945: 1944: 1908: 1902: 1901: 1891: 1876:Clin. Cancer Res 1867: 1861: 1860: 1850: 1818: 1812: 1811: 1801: 1791: 1767: 1761: 1760: 1750: 1740: 1716: 1707: 1706: 1696: 1672: 1666: 1665: 1650:Clin. Cancer Res 1645: 1639: 1638: 1628: 1618: 1586: 1580: 1579: 1553: 1536:(345): 345ra89. 1520: 1514: 1513: 1492: 1486: 1485: 1448: 1442: 1441: 1431: 1398: 1392: 1391: 1382:(4_suppl): 209. 1370: 1364: 1363: 1353: 1322:Physical Biology 1312: 1301: 1300: 1290: 1280: 1263:(8): 4589–4594. 1248: 1242: 1241: 1231: 1214:(6): 2206–2216. 1199: 1193: 1192: 1182: 1151:Physical Biology 1142: 1133: 1132: 1104: 1098: 1097: 1087: 1055: 1049: 1048: 1012: 1006: 1005: 969: 963: 962: 952: 928: 922: 921: 893: 887: 886: 876: 835: 829: 828: 796: 787: 786: 776: 751: 742: 741: 731: 699: 693: 692: 682: 658: 652: 651: 641: 609: 603: 602: 584: 560: 554: 553: 525: 382:Physical methods 333:Parsortix method 230:overall survival 4792: 4791: 4787: 4786: 4785: 4783: 4782: 4781: 4762: 4761: 4760: 4759: 4746:Design, ISITE. 4744: 4740: 4727: 4726: 4722: 4699:10.1038/nrc1098 4683: 4679: 4655: 4651: 4641: 4639: 4624: 4620: 4557: 4553: 4538: 4534: 4497: 4493: 4448: 4444: 4389: 4385: 4340: 4336: 4307:Clin Cancer Res 4299: 4295: 4258: 4254: 4223: 4219: 4163: 4159: 4113: 4109: 4086:10.1038/nm.3600 4074:Nature Medicine 4069: 4065: 4022: 4018: 3986: 3982: 3959:(12): 2787–95. 3945: 3941: 3901: 3897: 3850: 3846: 3823: 3819: 3772: 3768: 3721: 3717: 3670: 3666: 3621: 3617: 3572: 3568: 3531: 3527: 3487: 3483: 3457:10.1186/bcr1328 3450:(6): R975–979. 3436: 3432: 3393: 3389: 3373: 3372: 3322: 3318: 3286: 3282: 3274: 3267: 3263: 3262: 3258: 3229: 3225: 3220:Wayback Machine 3209: 3205: 3160: 3156: 3141: 3137: 3083: 3079: 3023: 3019: 2966: 2962: 2901: 2897: 2836: 2832: 2779: 2775: 2722: 2718: 2679: 2675: 2630: 2626: 2572: 2568: 2523: 2519: 2490:Clin Cancer Res 2482: 2478: 2449:Clin Cancer Res 2441: 2437: 2408:Clin Cancer Res 2400: 2396: 2373:(21): 6404–09. 2367:Clin Cancer Res 2359: 2355: 2310: 2306: 2297: 2295: 2285: 2281: 2280: 2276: 2267: 2265: 2256: 2255: 2251: 2220: 2216: 2171: 2167: 2127: 2118: 2073: 2069: 2038: 2034: 2003: 1999: 1952: 1948: 1909: 1905: 1868: 1864: 1833:(12): 3903–12. 1819: 1815: 1768: 1764: 1717: 1710: 1681:Am J Cancer Res 1673: 1669: 1646: 1642: 1601:(18): 4937–52. 1587: 1583: 1521: 1517: 1498:Cancer Research 1493: 1489: 1460:(11): 1531–41. 1449: 1445: 1399: 1395: 1371: 1367: 1313: 1304: 1249: 1245: 1200: 1196: 1143: 1136: 1109:Biotechnol. Adv 1105: 1101: 1070:(3–4): 101–14. 1056: 1052: 1013: 1009: 970: 966: 943:(12): 3646–50. 937:Clin Cancer Res 929: 925: 904:(18): 2645–50. 894: 890: 851:(7408): 510–3. 836: 832: 811:(19): 3213–21. 797: 790: 752: 745: 700: 696: 659: 655: 610: 606: 561: 557: 526: 522: 517: 487: 478: 476:Cell morphology 429: 400: 384: 375:Crohn's disease 358: 344: 335: 290: 242: 213: 205:carcinocythemia 165: 148: 92: 43:from a primary 17: 12: 11: 5: 4790: 4780: 4779: 4774: 4758: 4757: 4738: 4720: 4677: 4666:(7): 2073–84. 4649: 4618: 4551: 4532: 4491: 4462:(3): 132–142. 4442: 4383: 4334: 4313:(12): 3611–6. 4293: 4272:(5): 826–835. 4252: 4217: 4178:(25): 9393–8. 4157: 4107: 4080:(8): 897–903. 4063: 4036:(10): 667–72. 4016: 3980: 3939: 3895: 3844: 3833:(2): 427–442. 3817: 3766: 3715: 3664: 3615: 3586:(5): 821–828. 3566: 3525: 3498:(2): 211–218. 3481: 3430: 3403:(5): 597–605. 3387: 3316: 3280: 3277:on 2014-01-23. 3256: 3223: 3203: 3154: 3135: 3077: 3017: 2960: 2895: 2830: 2773: 2736:(4): 673–681. 2716: 2689:(3): 446–458. 2673: 2624: 2566: 2537:(3): 373–382. 2517: 2476: 2435: 2414:(19): 6302–9. 2394: 2353: 2304: 2274: 2249: 2230:(2): 180–204. 2214: 2165: 2132:J. Clin. Oncol 2116: 2087:(5): 528–538. 2067: 2032: 1997: 1946: 1903: 1882:(22): 6980–6. 1862: 1813: 1762: 1708: 1687:(6): 740–751. 1667: 1656:(8): 1950–60. 1640: 1581: 1515: 1487: 1443: 1414:(5): 1110–22. 1393: 1365: 1302: 1243: 1194: 1134: 1115:(7): 1063–84. 1099: 1050: 1007: 964: 923: 888: 830: 805:J. Clin. Oncol 788: 743: 714:(9): 1468–71. 694: 673:(3): 295–304. 653: 604: 555: 536:(1–2): 16–25. 519: 518: 516: 513: 486: 483: 477: 474: 428: 425: 399: 398:Hybrid methods 396: 383: 380: 360:Maintrac is a 357: 354: 343: 340: 334: 331: 289: 286: 241: 238: 212: 209: 164: 161: 147: 144: 143: 142: 138: 131: 119: 91: 88: 15: 9: 6: 4: 3: 2: 4789: 4778: 4775: 4773: 4770: 4769: 4767: 4753: 4749: 4742: 4734: 4730: 4724: 4716: 4712: 4708: 4704: 4700: 4696: 4692: 4688: 4681: 4673: 4669: 4665: 4661: 4653: 4642:September 25, 4637: 4633: 4629: 4622: 4614: 4610: 4605: 4600: 4596: 4592: 4587: 4582: 4578: 4574: 4570: 4566: 4562: 4555: 4547: 4543: 4536: 4528: 4524: 4519: 4514: 4511:(9): 1851–7. 4510: 4506: 4502: 4495: 4487: 4483: 4479: 4475: 4470: 4465: 4461: 4457: 4453: 4446: 4438: 4434: 4429: 4424: 4420: 4416: 4411: 4406: 4402: 4398: 4394: 4387: 4379: 4375: 4370: 4365: 4361: 4357: 4353: 4349: 4345: 4338: 4330: 4326: 4321: 4316: 4312: 4308: 4304: 4297: 4289: 4285: 4280: 4275: 4271: 4267: 4263: 4256: 4248: 4244: 4240: 4236: 4233:(5): 1111–8. 4232: 4228: 4221: 4213: 4209: 4204: 4199: 4194: 4189: 4185: 4181: 4177: 4173: 4169: 4161: 4153: 4149: 4144: 4139: 4135: 4131: 4127: 4123: 4119: 4111: 4103: 4099: 4095: 4091: 4087: 4083: 4079: 4075: 4067: 4059: 4055: 4051: 4047: 4043: 4039: 4035: 4031: 4027: 4020: 4012: 4008: 4004: 4000: 3996: 3992: 3984: 3976: 3972: 3967: 3962: 3958: 3954: 3950: 3943: 3935: 3931: 3927: 3923: 3919: 3915: 3912:(2): 355–64. 3911: 3907: 3899: 3891: 3887: 3882: 3877: 3872: 3867: 3863: 3859: 3855: 3848: 3840: 3836: 3832: 3828: 3821: 3813: 3809: 3804: 3799: 3794: 3789: 3785: 3781: 3777: 3770: 3762: 3758: 3753: 3748: 3743: 3738: 3734: 3730: 3726: 3719: 3711: 3707: 3702: 3697: 3692: 3687: 3683: 3679: 3675: 3668: 3660: 3656: 3651: 3646: 3642: 3638: 3634: 3630: 3626: 3619: 3611: 3607: 3602: 3597: 3593: 3589: 3585: 3581: 3577: 3570: 3562: 3558: 3553: 3548: 3544: 3540: 3536: 3529: 3521: 3517: 3513: 3509: 3505: 3501: 3497: 3493: 3485: 3477: 3473: 3468: 3463: 3458: 3453: 3449: 3445: 3441: 3434: 3426: 3422: 3418: 3414: 3410: 3406: 3402: 3398: 3391: 3383: 3377: 3369: 3365: 3360: 3355: 3351: 3347: 3343: 3339: 3336:(1): 016002. 3335: 3331: 3327: 3320: 3312: 3308: 3304: 3300: 3296: 3292: 3284: 3273: 3266: 3260: 3251: 3246: 3242: 3238: 3234: 3227: 3221: 3217: 3214: 3207: 3199: 3195: 3190: 3185: 3181: 3177: 3173: 3169: 3165: 3158: 3150: 3149:www.gilupi.de 3146: 3139: 3131: 3127: 3122: 3117: 3113: 3109: 3105: 3101: 3097: 3093: 3089: 3081: 3073: 3069: 3064: 3059: 3054: 3049: 3045: 3041: 3038:(9): e71105. 3037: 3033: 3029: 3021: 3013: 3009: 3004: 2999: 2995: 2991: 2987: 2983: 2979: 2975: 2971: 2964: 2956: 2952: 2947: 2942: 2938: 2934: 2930: 2926: 2922: 2918: 2914: 2910: 2906: 2899: 2891: 2887: 2882: 2877: 2873: 2869: 2865: 2861: 2857: 2853: 2849: 2845: 2841: 2834: 2826: 2822: 2817: 2812: 2808: 2804: 2800: 2796: 2792: 2788: 2784: 2777: 2769: 2765: 2760: 2755: 2751: 2747: 2743: 2739: 2735: 2731: 2727: 2720: 2712: 2708: 2704: 2700: 2696: 2692: 2688: 2684: 2683:Lab on a Chip 2677: 2669: 2665: 2660: 2655: 2651: 2647: 2643: 2639: 2638:Lab on a Chip 2635: 2628: 2620: 2616: 2611: 2606: 2602: 2598: 2594: 2590: 2586: 2582: 2578: 2570: 2562: 2558: 2553: 2548: 2544: 2540: 2536: 2532: 2528: 2521: 2513: 2509: 2504: 2499: 2495: 2491: 2487: 2480: 2472: 2468: 2463: 2458: 2454: 2450: 2446: 2439: 2431: 2427: 2422: 2417: 2413: 2409: 2405: 2398: 2390: 2386: 2381: 2376: 2372: 2368: 2364: 2357: 2349: 2345: 2341: 2337: 2332: 2327: 2324:(8): 781–91. 2323: 2319: 2315: 2308: 2293: 2292: 2284: 2278: 2264:on 2008-06-05 2263: 2259: 2253: 2245: 2241: 2237: 2233: 2229: 2225: 2218: 2210: 2206: 2202: 2198: 2193: 2188: 2184: 2180: 2176: 2169: 2161: 2157: 2153: 2149: 2145: 2141: 2137: 2133: 2125: 2123: 2121: 2112: 2108: 2103: 2098: 2094: 2090: 2086: 2082: 2081:Transl. Oncol 2078: 2071: 2063: 2059: 2055: 2051: 2047: 2043: 2036: 2028: 2024: 2020: 2016: 2012: 2008: 2001: 1993: 1989: 1984: 1979: 1974: 1969: 1965: 1961: 1957: 1950: 1942: 1938: 1934: 1930: 1926: 1922: 1919:(1–2): 1–13. 1918: 1914: 1907: 1899: 1895: 1890: 1885: 1881: 1877: 1873: 1866: 1858: 1854: 1849: 1844: 1840: 1836: 1832: 1828: 1824: 1817: 1809: 1805: 1800: 1795: 1790: 1785: 1781: 1777: 1773: 1766: 1758: 1754: 1749: 1744: 1739: 1734: 1730: 1726: 1722: 1715: 1713: 1704: 1700: 1695: 1690: 1686: 1682: 1678: 1671: 1663: 1659: 1655: 1651: 1644: 1636: 1632: 1627: 1622: 1617: 1612: 1608: 1604: 1600: 1596: 1592: 1585: 1577: 1573: 1569: 1565: 1561: 1557: 1552: 1547: 1543: 1539: 1535: 1531: 1527: 1519: 1511: 1507: 1503: 1499: 1491: 1483: 1479: 1475: 1471: 1467: 1463: 1459: 1455: 1447: 1439: 1435: 1430: 1425: 1421: 1417: 1413: 1409: 1405: 1397: 1389: 1385: 1381: 1377: 1369: 1361: 1357: 1352: 1347: 1343: 1339: 1335: 1331: 1328:(1): 016003. 1327: 1323: 1319: 1311: 1309: 1307: 1298: 1294: 1289: 1284: 1279: 1274: 1270: 1266: 1262: 1258: 1254: 1247: 1239: 1235: 1230: 1225: 1221: 1217: 1213: 1209: 1205: 1198: 1190: 1186: 1181: 1176: 1172: 1168: 1164: 1160: 1157:(1): 016004. 1156: 1152: 1148: 1141: 1139: 1130: 1126: 1122: 1118: 1114: 1110: 1103: 1095: 1091: 1086: 1081: 1077: 1073: 1069: 1065: 1061: 1054: 1046: 1042: 1038: 1034: 1030: 1026: 1022: 1018: 1011: 1003: 999: 995: 991: 987: 983: 980:(6): 339–51. 979: 975: 968: 960: 956: 951: 946: 942: 938: 934: 927: 919: 915: 911: 907: 903: 899: 892: 884: 880: 875: 870: 866: 862: 858: 854: 850: 846: 842: 834: 826: 822: 818: 814: 810: 806: 802: 795: 793: 784: 780: 775: 770: 767:(7): 2912–8. 766: 762: 758: 750: 748: 739: 735: 730: 725: 721: 717: 713: 709: 705: 698: 690: 686: 681: 676: 672: 668: 664: 657: 649: 645: 640: 635: 631: 627: 623: 619: 615: 608: 600: 596: 592: 588: 583: 578: 575:(4): 679–95. 574: 570: 566: 559: 551: 547: 543: 539: 535: 531: 524: 520: 512: 510: 504: 502: 496: 493: 482: 473: 470: 465: 463: 460:status using 459: 455: 451: 447: 443: 438: 434: 424: 422: 418: 414: 410: 406: 395: 393: 389: 379: 376: 370: 367: 363: 353: 350: 339: 330: 328: 324: 320: 316: 313: 309: 304: 299: 295: 294:nanoparticles 285: 283: 279: 275: 271: 267: 263: 259: 255: 251: 247: 237: 233: 231: 227: 217: 208: 206: 202: 196: 194: 190: 186: 181: 177: 169: 160: 156: 152: 139: 136: 132: 128: 124: 120: 117: 113: 109: 105: 104: 103: 101: 97: 87: 85: 81: 80:liquid biopsy 76: 74: 70: 66: 62: 58: 54: 50: 46: 42: 38: 34: 30: 21: 4751: 4741: 4732: 4723: 4693:(6): 453–8. 4690: 4686: 4680: 4663: 4659: 4652: 4640:. 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