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Isolation Of Rare Circulating Tumour Cells In Cancer Patients

7750 words - 31 pages

Vol 450 | 20/27 December 2007 | doi:10.1038/nature06385

LETTERS
Isolation of rare circulating tumour cells in cancer patients by microchip technology
Sunitha Nagrath1*, Lecia V. Sequist2*, Shyamala Maheswaran2, Daphne W. Bell2{, Daniel Irimia1, Lindsey Ulkus2, Matthew R. Smith2, Eunice L. Kwak2, Subba Digumarthy2, Alona Muzikansky2, Paula Ryan2, Ulysses J. Balis1{, Ronald G. Tompkins1, Daniel A. Haber2 & Mehmet Toner1

Viable tumour-derived epithelial cells (circulating tumour cells or CTCs) have been identified in peripheral blood from cancer patients and are probably the origin of intractable metastatic disease1–4. Although extremely rare, CTCs represent a potential alternative to ...view middle of the document...

In a small cohort of patients with metastatic cancer undergoing systemic treatment, temporal changes in CTC numbers correlated reasonably well with the clinical course of disease as measured by standard radiographic methods. Thus, the CTC-chip provides a new and effective tool for accurate identification and measurement of CTCs in patients with cancer. It has broad implications in advancing both cancer biology research and clinical cancer management, including the detection, diagnosis and monitoring of cancer10. CTCs are rare, comprising as few as one cell per 109 haematologic cells in the blood of patients with metastatic cancer, hence their isolation presents a tremendous technical challenge7,9,11–13. Microfluidic lab-on-a-chip devices provide unique opportunities for cell sorting and rare-cell detection; they have been successfully used for microfluidic flow cytometry14, continuous size-based separation15,16 and chromatographic separation17. Despite their success in manipulating microlitre amounts of simple liquids in microscale channels14,18,19, they have thus far shown limited capability to deal with the cellular and fluid complexity of large volumes (millilitres) of whole blood samples20–22.
1 2

Here we describe the development and application of a microfluidic device (the ‘CTC-chip’) that can efficiently and reproducibly isolate CTCs from the blood of patients with common epithelial tumours (Fig. 1, and Supplementary Fig. 1). The CTC-chip (Fig. 1b) consists of an array of microposts (Supplementary Fig. 1c) that are made chemically functional with anti-epithelial-celladhesion-molecule (EpCAM, also known as TACSTD1) antibodies. Anti-EpCAM provides the specificity for CTC capture from unfractionated blood because EpCAM is frequently overexpressed by carcinomas of lung, colorectal, breast, prostate, head and neck, and hepatic origin, and is absent from haematologic cells23,24. Two essential parameters that determine the efficiency of cell capture on the CTC-chip are: (1) flow velocity, because it influences the duration of cell–micropost contact; and (2) shear force, which must be sufficiently low to ensure maximum cell–micropost attachment. To optimize these parameters we employed theoretical analyses characterizing the interaction of cells with microposts distributed within
a
Pressure control Whole blood Rocker assembly

c

Manifold housing CTC-chip

d

Pressure source

b

Figure 1 | Isolation of CTCs from whole blood using a microfluidic device. a, The workstation setup for CTC separation. The sample is continually mixed on a rocker, and pumped through the chip using a pneumaticpressure-regulated pump. b, The CTC-chip with microposts etched in silicon. c, Whole blood flowing through the microfluidic device. d, Scanning electron microscope image of a captured NCI-H1650 lung cancer cell spiked into blood (pseudo coloured red). The inset shows a high magnification view of the cell.

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