Circulating tumor cell microseparator based on lateral magnetophoresis and immunomagnetic nanobeads.

This paper presents a circulating tumor cell (CTC) microseparator for isolation of CTCs from human peripheral blood using immunomagnetic nanobeads with bound antiepithelial cell adhesive molecule (EpCAM) antibodies that specifically bind to epithelial cancer cells. The isolation is performed through lateral magnetophoresis, which is induced by high-gradient magnetic separation technology, involving a ferromagnetic wire array inlaid in the bottom substrate of a microchannel. Experimental results showed that the CTC microseparator isolates about 90% of spiked CTCs in human peripheral blood at a flow rate of up to 5 mL/h and purifies to approximately 97%. The overall isolation procedure was completed within 15 min for 200 μL of peripheral blood. CTCs from peripheral blood of patients with breast and lung cancers were isolated with the CTC microseparator, and the results were compared with those of healthy donors. Using a fluorescence-based viability assay, the viability of CTCs isolated from peripheral blood of patients with cancer was observed. In addition, the usefulness of the CTC microseparator for subsequent genetic assay was confirmed by reverse-transcriptase polymerase chain reaction (RT-PCR) amplification of cancer-specific genes using CTCs isolated from patients with cancer.

[1]  Han,et al.  Paramagnetic capture mode magnetophoretic microseparator for blood cells , 2005 .

[2]  Katharina Pachmann,et al.  Standardized quantification of circulating peripheral tumor cells from lung and breast cancer , 2005, Clinical chemistry and laboratory medicine.

[3]  Kazunori Hoshino,et al.  Microchip-based immunomagnetic detection of circulating tumor cells. , 2011, Lab on a chip.

[4]  S. Groshen,et al.  Portable Filter-Based Microdevice for Detection and Characterization of Circulating Tumor Cells , 2010, Clinical Cancer Research.

[5]  P. V. van Dam,et al.  Circulating tumour cell detection: a direct comparison between the CellSearch System, the AdnaTest and CK-19/mammaglobin RT–PCR in patients with metastatic breast cancer , 2009, British Journal of Cancer.

[6]  J. Moldvay,et al.  The role of TTF-1 in differentiating primary and metastatic lung adenocarcinomas , 2008, Pathology & Oncology Research.

[7]  Paul I. Okagbare,et al.  Highly efficient circulating tumor cell isolation from whole blood and label-free enumeration using polymer-based microfluidics with an integrated conductivity sensor. , 2008, Journal of the American Chemical Society.

[8]  M C Miller,et al.  Expression of epithelial cell adhesion molecule in carcinoma cells present in blood and primary and metastatic tumors. , 2004, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[9]  C. Rao,et al.  Optimization of ferrofluids and protocols for the enrichment of breast tumor cells in blood , 2001 .

[10]  Peter Kuhn,et al.  High speed detection of circulating tumor cells. , 2006, Biosensors & bioelectronics.

[11]  J R Siewert,et al.  Comparison of two density gradient centrifugation systems for the enrichment of disseminated tumor cells in blood. , 2002, Cytometry.

[12]  Jonathan W. Uhr,et al.  Tumor Cells Circulate in the Peripheral Blood of All Major Carcinomas but not in Healthy Subjects or Patients With Nonmalignant Diseases , 2004, Clinical Cancer Research.

[13]  Mehmet Toner,et al.  The CTC-chip: an exciting new tool to detect circulating tumor cells in lung cancer patients. , 2009, Journal of thoracic oncology : official publication of the International Association for the Study of Lung Cancer.

[14]  Bo Lu,et al.  3D microfilter device for viable circulating tumor cell (CTC) enrichment from blood , 2011, Biomedical microdevices.

[15]  Tanja Fehm,et al.  Cytogenetic evidence that circulating epithelial cells in patients with carcinoma are malignant. , 2002, Clinical cancer research : an official journal of the American Association for Cancer Research.

[16]  Unyoung Kim,et al.  Multitarget magnetic activated cell sorter , 2008, Proceedings of the National Academy of Sciences.

[17]  Robert H. Austin,et al.  Continuous microfluidic immunomagnetic cell separation , 2004 .

[18]  R. Eils,et al.  Systemic spread is an early step in breast cancer. , 2008, Cancer cell.

[19]  S. Digumarthy,et al.  Isolation of rare circulating tumour cells in cancer patients by microchip technology , 2007, Nature.

[20]  M. Dowsett,et al.  Detection of circulating epithelial cells in the blood of patients with breast cancer: comparison of three techniques , 2005, British Journal of Cancer.

[21]  K. Isselbacher,et al.  Isolation of circulating tumor cells using a microvortex-generating herringbone-chip , 2010, Proceedings of the National Academy of Sciences.

[22]  Ruud H. Brakenhoff,et al.  Detection, clinical relevance and specific biological properties of disseminating tumour cells , 2008, Nature Reviews Cancer.

[23]  G. Doyle,et al.  Changes in circulating carcinoma cells in patients with metastatic prostate cancer correlate with disease status. , 2001, Urology.

[24]  A. Bruno Frazier,et al.  Continuous magnetophoretic separation of blood cells in microdevice format , 2004 .

[25]  G. Assmann,et al.  An immunological enrichment method for epithelial cells from peripheral blood. , 1995, Journal of immunological methods.

[26]  Bo Lu,et al.  A cancer detection platform which measures telomerase activity from live circulating tumor cells captured on a microfilter. , 2010, Cancer research.

[27]  J. W. Parce,et al.  Electrokinetically controlled microfluidic analysis systems. , 2000, Annual review of biophysics and biomolecular structure.

[28]  Peter Kuhn,et al.  A rare-cell detector for cancer. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[29]  H Heynemann,et al.  Detection and enrichment of disseminated renal carcinoma cells from peripheral blood by immunomagnetic cell separation , 2001, International journal of cancer.

[30]  Hwanyong Lee,et al.  High-speed RNA microextraction technology using magnetic oligo-dT beads and lateral magnetophoresis. , 2010, Lab on a chip.

[31]  A. Weiss,et al.  Detection and characterization of carcinoma cells in the blood. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[32]  K. Schütze,et al.  Isolation by size of epithelial tumor cells : a new method for the immunomorphological and molecular characterization of circulatingtumor cells. , 2000, The American journal of pathology.

[33]  Alison Stopeck,et al.  Circulating tumor cells, disease progression, and survival in metastatic breast cancer. , 2004, The New England journal of medicine.

[34]  Thomas Rau,et al.  Circulating Tumor Cells in Breast Cancer: Correlation to Bone Marrow Micrometastases, Heterogeneous Response to Systemic Therapy and Low Proliferative Activity , 2005, Clinical Cancer Research.

[35]  M. Naoe,et al.  Detection of circulating urothelial cancer cells in the blood using the CellSearch System , 2007, Cancer.