Challenges in circulating tumour cell research

Alix‐Panabières, Catherine; Pantel, Klaus

doi:10.1038/nrc3820

Cited by 1,136 publications

(1,023 citation statements)

References 96 publications

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“…However, many cancers, especially those originating in the epithelial tissues, are known to modulate the expression of surface proteins, making acquisition difficult due to changing surface antigens. 4,5 In many cases, this issue compounds application of flow cytometric and magnetic sorting by requiring multiple expensive antibodies to target cells with sufficient accuracy and certainty. Stress caused to the cells via the use of several compounds may alter the behavior and phenotype of the cell.…”

Section: Introductionmentioning

confidence: 99%

Enhanced contactless dielectrophoresis enrichment and isolation platform via cell-scale microstructures

et al. 2016

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We designed a new microfluidic device that uses pillars on the same order as the diameter of a cell (20 lm) to isolate and enrich rare cell samples from background. These cell-scale microstructures improve viability, trapping efficiency, and throughput while reducing pearl chaining. The area where cells trap on each pillar is small, such that only one or two cells trap while fluid flow carries away excess cells. We employed contactless dielectrophoresis in which a thin PDMS membrane separates the cell suspension from the electrodes, improving cell viability for off-chip collection and analysis. We compared viability and trapping efficiency of a highly aggressive Mouse Ovarian Surface Epithelial (MOSE) cell line in this 20 lm pillar device to measurements in an earlier device with the same layout but pillars of 100 lm diameter. We found that MOSE cells in the new device with 20 lm pillars had higher viability at 350 V RMS , 30 kHz, and 1.2 ml/h (control 77%, untrapped 71%, trapped 81%) than in the previous generation device (untrapped 47%, trapped 42%). The new device can trap up to 6 times more cells under the same conditions. Our new device can sort cells with a high flow rate of 2.2 ml/h and throughput of a few million cells per hour while maintaining a viable population of cells for off-chip analysis. By using the device to separate subpopulations of tumor cells while maintaining their viability at large sample sizes, this technology can be used in developing personalized treatments that target the most aggressive cancerous cells. V C 2016 AIP Publishing LLC. [http://dx

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Section: Introductionmentioning

confidence: 99%

Enhanced contactless dielectrophoresis enrichment and isolation platform via cell-scale microstructures

et al. 2016

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“…Biontinylated horse anti-goat antibody (Vector Labs BA-9500, 10 μg/mL) was incubated in the streptavidin-coated sites for 30 minutes, followed by rinsing for 5 times with 1 × phosphate-buffered saline (PBS). Then, goat anti-EpCAM antibody (RnD AF960, 10 μg/mL) was further incubated only in the target wells for 30 minutes and followed by rinsing for 5 times with 1 × PBS, such that the target wells can capture MCF-7, which has epithelial cell adhesion molecules (EpCAM) as surface markers [3]. 10 μL MCF-7 in 1 × PBS was loaded to all the wells on the substrate for 30 minutes that allowed binding between the antibodies and EpCAM, and thus capture of MCF-7 (Fig.…”

Section: Sample Preparationsmentioning

confidence: 99%

“…In this example, we utilized EpCAM, a cell adhesion molecule commonly expressed on epithelial cells, as a biomarker to distinguish MCF-7 cells from the white blood cells through an immunological binding approach. Therefore, our system not only can perform EpCAM-based CTC enrichment, similar to the existing techniques [3,21,22], but also allows in situ quantitative image analysis of the captured cells with the single-cell precision, thanks to the high-resolution and high-throughput imaging capability. Notably, coupled with quantitative phase time-stretch imaging, this high-throughput spinning imaging cell-based assay could allow single-cell biophysical phenotyping, e.g.…”

Section: Cancer Cell Screening: Mixture Of Human Blood and Mcf-7mentioning

confidence: 99%

“…However, this throughput is still at least two-orders-of-magnitude slower than classical non-imaging flow cytometry because of the inherent speed-versus-sensitivity limitation of the camera technologies. This pervasive problem hinders the acceptance of imaging cell-based assay in high-throughput screening (HTS) applications [3][4][5], e.g. phenotypic screening in the early drug discovery pipeline in which tens of thousands compounds per screen are involved; and circulating tumor cell (CTC) screening in which rare cell detection within enormous and heterogeneous population (some millions of blood cells) is mandated.…”

Section: Introductionmentioning

confidence: 99%

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Time-stretch microscopy on a DVD for high-throughput imaging cell-based assay

Tang¹,

Yeung²,

Chan³

et al. 2017

Biomed. Opt. Express

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Cell-based assay based on time-stretch imaging is recognized to be well-suited for high-throughput phenotypic screening. However, this ultrafast imaging technique has primarily been limited to suspension-cell assay, leaving a wide range of solid-substrate assay formats uncharted. Moreover, time-stretch imaging is generally restricted to intrinsic biophysical phenotyping, but lacks the biomolecular signatures of the cells. To address these challenges, we develop a spinning time-stretch imaging assay platform based on the functionalized digital versatile disc (DVD). We demonstrate that adherent cell culture and biochemically-specific cell-capture can now be assayed with time-stretch microscopy, thanks to the high-speed DVD spinning motion that naturally enables on-the-fly cellular imaging at an ultrafast line-scan rate of >10MHz. As scanning the whole DVD at such a high speed enables ultra-large field-of-view imaging, it could be favorable for scaling both the assay throughput and content as demanded in many applications, e.g. drug discovery, and rare cancer cell screening. Balakrishnan, C.-Y. Wang, P. Yaswen, A. Goga, and Z. Werb, "Single-cell analysis reveals a stem-cell program in human metastatic breast cancer cells," Nature 526(7571), 131-135 (2015).

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“…A variety of technologies have been developed to enrich CTCs that commonly rely on cell surface antigen expression or on physical properties including cell size and deformability that distinguish CTCs from blood cells (Alix‐Panabières and Pantel, 2014; Alix‐Panabières and Pantel, 2016; Chudziak et al ., 2016; Ferreira et al ., 2016; Pantel, 2016). CellSearch ® is a FDA‐approved epitope‐dependent platform capable of CTC enrichment and enumeration.…”

Section: Introductionmentioning

confidence: 99%

Molecular analysis of single circulating tumour cells following long‐term storage of clinical samples

et al. 2017

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The CellSearch® semiautomated CTC enrichment and staining system has been established as the ‘gold standard’ for CTC enumeration with CellSearch® CTC counts recognized by the FDA as prognostic for a number of cancers. We and others have gone on to show that molecular analysis of CellSearch® CTCs isolated shortly after CellSearch® enrichment provides another valuable layer of information that has potential clinical utility including predicting response to treatment. Although CellSearch® CTCs can be readily isolated after enrichment, the process of analysing a single CellSearch® patient sample, which may contain many CTCs, is both time‐consuming and costly. Here, we describe a simple process that will allow storage of all CellSearch®‐enriched cells in glycerol at −20 °C for up to 2 years without any measurable loss in the ability to retrieve single cells or in the genome integrity of the isolated cells. To establish the suitability of long‐term glycerol storage for single‐cell molecular analysis, we isolated individual CellSearch®‐enriched cells by DEPArray™ either shortly after CellSearch® enrichment or following storage of matched enriched cells in glycerol at −20 °C. All isolated cells were subjected to whole‐genome amplification (WGA), and the efficacy of single‐cell WGA was evaluated by multiplex PCR to generate a Genome Integrity Index (GII). The GII results from 409 single cells obtained from both ‘spike‐in’ controls and clinical samples showed no statistical difference between values obtained pre‐ and postglycerol storage and that there is no further loss in integrity when DEPArray™‐isolated cells are then stored at −80 °C for up to 2 years. In summary, we have established simple yet effective ‘stop‐off’ points along the CTC workflow enabling CTC banking and facilitating selection of suitable samples for intensive analysis once patient outcomes are known.

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Challenges in circulating tumour cell research

Cited by 1,136 publications

References 96 publications

Enhanced contactless dielectrophoresis enrichment and isolation platform via cell-scale microstructures

Enhanced contactless dielectrophoresis enrichment and isolation platform via cell-scale microstructures

Time-stretch microscopy on a DVD for high-throughput imaging cell-based assay

Molecular analysis of single circulating tumour cells following long‐term storage of clinical samples

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