Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

Isolation of rare circulating tumour cells in cancer patients by microchip technology

Abstract

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 disease1,2,3,4. Although extremely rare, CTCs represent a potential alternative to invasive biopsies as a source of tumour tissue for the detection, characterization and monitoring of non-haematologic cancers5,6,7,8. The ability to identify, isolate, propagate and molecularly characterize CTC subpopulations could further the discovery of cancer stem cell biomarkers and expand the understanding of the biology of metastasis. Current strategies for isolating CTCs are limited to complex analytic approaches that generate very low yield and purity9. Here we describe the development of a unique microfluidic platform (the ‘CTC-chip’) capable of efficient and selective separation of viable CTCs from peripheral whole blood samples, mediated by the interaction of target CTCs with antibody (EpCAM)-coated microposts under precisely controlled laminar flow conditions, and without requisite pre-labelling or processing of samples. The CTC-chip successfully identified CTCs in the peripheral blood of patients with metastatic lung, prostate, pancreatic, breast and colon cancer in 115 of 116 (99%) samples, with a range of 5–1,281 CTCs per ml and approximately 50% purity. In addition, CTCs were isolated in 7/7 patients with early-stage prostate cancer. Given the high sensitivity and specificity of the CTC-chip, we tested its potential utility in monitoring response to anti-cancer therapy. 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.

Access options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

Figure 1: Isolation of CTCs from whole blood using a microfluidic device.
Figure 2: CTC capture and enumeration.
Figure 3: Enumeration of CTCs from cancer patients.
Figure 4: Characterization of CTCs with tumour-specific molecular markers.

References

  1. 1

    Cristofanilli, M. et al. Circulating tumor cells, disease progression, and survival in metastatic breast cancer. N. Engl. J. Med. 351, 781–791 (2004)

    CAS  Article  Google Scholar 

  2. 2

    Steeg, P. S. Tumor metastasis: mechanistic insights and clinical challenges. Nature Med. 12, 895–904 (2006)

    CAS  Article  Google Scholar 

  3. 3

    Gupta, G. P. & Massague, J. Cancer metastasis: building a framework. Cell 127, 679–695 (2006)

    CAS  Article  Google Scholar 

  4. 4

    Reya, T., Morrison, S. J., Clarke, M. F. & Weissman, I. L. Stem cells, cancer, and cancer stem cells. Nature 414, 105–111 (2001)

    ADS  CAS  Article  Google Scholar 

  5. 5

    Mocellin, S., Hoon, D., Ambrosi, A., Nitti, D. & Rossi, C. R. The prognostic value of circulating tumor cells in patients with melanoma: a systematic review and meta-analysis. Clin. Cancer Res. 12, 4605–4613 (2006)

    CAS  Article  Google Scholar 

  6. 6

    Smerage, J. B. & Hayes, D. F. The measurement and therapeutic implications of circulating tumour cells in breast cancer. Br. J. Cancer 94, 8–12 (2006)

    CAS  Article  Google Scholar 

  7. 7

    Rolle, A. et al. Increase in number of circulating disseminated epithelial cells after surgery for non-small cell lung cancer monitored by MAINTRAC(R) is a predictor for relapse: A preliminary report. World J. Surg. Oncol. 3, 18 (2005)

    ADS  Article  Google Scholar 

  8. 8

    Braun, S. & Marth, C. Circulating tumor cells in metastatic breast cancer—toward individualized treatment? N. Engl. J. Med. 351, 824–826 (2004)

    CAS  Article  Google Scholar 

  9. 9

    Zieglschmid, V., Hollmann, C. & Bocher, O. Detection of disseminated tumor cells in peripheral blood. Crit. Rev. Clin. Lab. Sci. 42, 155–196 (2005)

    CAS  Article  Google Scholar 

  10. 10

    Bell, D. W. & Haber, D. A. A blood-based test for epidermal growth factor receptor mutations in lung cancer. Clin. Cancer Res. 12, 3875–3877 (2006)

    CAS  Article  Google Scholar 

  11. 11

    Kahn, H. J. et al. Enumeration of circulating tumor cells in the blood of breast cancer patients after filtration enrichment: correlation with disease stage. Breast Cancer Res. Treat. 86, 237–247 (2004)

    Article  Google Scholar 

  12. 12

    Krivacic, R. T. et al. A rare-cell detector for cancer. Proc. Natl Acad. Sci. USA 101, 10501–10504 (2004)

    ADS  CAS  Article  Google Scholar 

  13. 13

    Racila, E. et al. Detection and characterization of carcinoma cells in the blood. Proc. Natl Acad. Sci. USA 95, 4589–4594 (1998)

    ADS  CAS  Article  Google Scholar 

  14. 14

    Fu, A. Y., Spence, C., Scherer, A., Arnold, F. H. & Quake, S. R. A microfabricated fluorescence-activated cell sorter. Nature Biotechnol. 17, 1109–1111 (1999)

    CAS  Article  Google Scholar 

  15. 15

    Davis, J. A. et al. Deterministic hydrodynamics: taking blood apart. Proc. Natl Acad. Sci. USA 103, 14779–14784 (2006)

    ADS  CAS  Article  Google Scholar 

  16. 16

    Huang, L. R., Cox, E. C., Austin, R. H. & Sturm, J. C. Continuous particle separation through deterministic lateral displacement. Science 304, 987–990 (2004)

    ADS  CAS  Article  Google Scholar 

  17. 17

    Chang, W. C., Lee, L. P. & Liepmann, D. Biomimetic technique for adhesion-based collection and separation of cells in a microfluidic channel. Lab Chip 5, 64–73 (2005)

    CAS  Article  Google Scholar 

  18. 18

    Whitesides, G. M. The origins and the future of microfluidics. Nature 442, 368–373 (2006)

    ADS  CAS  Article  Google Scholar 

  19. 19

    Hong, J. W. & Quake, S. R. Integrated nanoliter systems. Nature Biotechnol. 21, 1179–1183 (2003)

    CAS  Article  Google Scholar 

  20. 20

    Toner, M. & Irimia, D. Blood-on-a-chip. Annu. Rev. Biomed. Eng. 7, 77–103 (2005)

    CAS  Article  Google Scholar 

  21. 21

    Dittrich, P. S. & Manz, A. Lab-on-a-chip: microfluidics in drug discovery. Nature Rev. Drug Discov. 5, 210–218 (2006)

    CAS  Article  Google Scholar 

  22. 22

    El-Ali, J., Sorger, P. K. & Jensen, K. F. Cells on chips. Nature 442, 403–411 (2006)

    ADS  CAS  Article  Google Scholar 

  23. 23

    Went, P. T. et al. Frequent EpCam protein expression in human carcinomas. Hum. Pathol. 35, 122–128 (2004)

    CAS  Article  Google Scholar 

  24. 24

    Balzar, M., Winter, M. J., de Boer, C. J. & Litvinov, S. V. The biology of the 17–1A antigen (Ep-CAM). J. Mol. Med. 77, 699–712 (1999)

    CAS  Article  Google Scholar 

  25. 25

    Rao, C. G. et al. Expression of epithelial cell adhesion molecule in carcinoma cells present in blood and primary and metastatic tumors. Int. J. Oncol. 27, 49–57 (2005)

    CAS  PubMed  Google Scholar 

  26. 26

    Smirnov, D. A. et al. Global gene expression profiling of circulating tumor cells. Cancer Res. 65, 4993–4997 (2005)

    CAS  Article  Google Scholar 

  27. 27

    Therasse, P. et al. New guidelines to evaluate the response to treatment in solid tumors. European Organization for Research and Treatment of Cancer, National Cancer Institute of the United States, National Cancer Institute of Canada. J. Natl. Cancer Inst. 92, 205–216 (2000)

    CAS  Article  Google Scholar 

  28. 28

    Terstappen, L. W. et al. Flow cytometry–principles and feasibility in transfusion medicine. Enumeration of epithelial derived tumor cells in peripheral blood. Vox Sang. 74 (suppl. 2). 269–274 (1998)

    CAS  Article  Google Scholar 

  29. 29

    Kraeft, S. K. et al. Reliable and sensitive identification of occult tumor cells using the improved rare event imaging system. Clin. Cancer Res. 10, 3020–3028 (2004)

    CAS  Article  Google Scholar 

  30. 30

    Allard, W. J. et al. Tumor cells circulate in the peripheral blood of all major carcinomas but not in healthy subjects or patients with nonmalignant diseases. Clin. Cancer Res. 10, 6897–6904 (2004)

    Article  Google Scholar 

  31. 31

    Drummond, J. E. & Tahir, M. I. Laminar viscous flow through regular arrays of parallel solid cylinders. Int. J. Multiphase Flow 10, 515–539 (1984)

    CAS  Article  Google Scholar 

  32. 32

    Murthy, S. K., Sin, A., Tompkins, R. G. & Toner, M. Effect of flow and surface conditions on human lymphocyte isolation using microfluidic chambers. Langmuir 20, 11649–11655 (2004)

    CAS  Article  Google Scholar 

Download references

Acknowledgements

We thank A. Amin for technical assistance in running experiments, O. Hurtado for clean room work, S. Murthy for surface chemistry, L. Romonosky for cell counting, D. Hyde for digital pictures and D. Poulsen for illustrations. We are also grateful to R. Kapur and his team for technical assistance. The authors acknowledge funding from the National Institutes of Health (to M.T.), and the Doris Duke Distinguished Clinical Scientist Award (to D.A.H.).

Author Contributions S.N., L.V.S., R.G.T., D.A.H. and M.T. designed and conducted the study. S.M., D.W.B. and L.U. performed gene expression analyses; D.I. contributed to the microfluidic system. M.R.S., E.L.K. and P.R. acquired clinical samples. U.J.B. provided input on cytopathology; A.M. performed statistical analysis; and S.D. performed radiology measurements. S.N., L.V.S., D.W.B., S.M., D.I., D.A.H. and M.T. participated in data analysis and writing of the manuscript.

Author information

Affiliations

Authors

Corresponding author

Correspondence to Mehmet Toner.

Ethics declarations

Competing interests

At the time the reported study was performed, M.T., R.G.T. and Massachusetts General Hospital (MGH) had significant equity holdings or similar interests in licensees from MGH for technology described in this manuscript. Before final submission for publication, recognizing Partners HealthCare System and Harvard Medical School policies on conflicts of interest, M.T., R.G.T. and MGH relinquished all such interests.

Supplementary information

Supplementary Information

The file contains Supplementary Figures S1-S6 with Legends and Supplementary Tables S1-S2. (PDF 852 kb)

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Nagrath, S., Sequist, L., Maheswaran, S. et al. Isolation of rare circulating tumour cells in cancer patients by microchip technology. Nature 450, 1235–1239 (2007). https://doi.org/10.1038/nature06385

Download citation

Further reading

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.

Search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing