Rare cell isolation and recovery on open-channel microfluidic chip

The ability to accurately detect and analyze rare cells in a cell population is critical not only for the study of disease progression but also for next flow cytometry systems in clinical application. Here, we report the development of a prototype device, the ‘Rare cell sorter’, for isolating and recovering single rare cells from whole blood samples. On this device, we utilized an open-channel microfluidic chip for rare cell isolation. And the advantage of open-channel allows us to recover the isolated rare cell directly from the chip. We set the circulating tumor cell (CTC) as a target cell. For the clinical experiment, CTCs were isolated from blood samples collected from patients with metastatic breast cancer and healthy volunteers. There was a significant difference in the number of CTCs between the patients with metastatic breast cancer and healthy volunteers. To evaluate the damage to cells during isolation and recovery, we performed an RNA integrity assay using RNA extracted from CTCs recovered from the chip and found that our process for single CTC isolation and recovery is mild enough for gene analysis of CTCs.

[1]  Mina J Bissell,et al.  Isolation, immortalization, and characterization of a human breast epithelial cell line with stem cell properties. , 2002, Genes & development.

[2]  Brian A. Smith,et al.  Viable circulating metastatic cells produced in orthotopic but not ectopic prostate cancer models. , 2003, Cancer research.

[3]  T. Kraus,et al.  Role of the meniscus shape in large-area convective particle assembly. , 2011, Langmuir : the ACS journal of surfaces and colloids.

[4]  M. Lidstrom,et al.  The role of physiological heterogeneity in microbial population behavior. , 2010, Nature chemical biology.

[5]  Sridhar Ramaswamy,et al.  A microfluidic device for label-free, physical capture of circulating tumor cell-clusters , 2015, Nature Methods.

[6]  Siyang Zheng,et al.  Membrane microfilter device for selective capture, electrolysis and genomic analysis of human circulating tumor cells. , 2007, Journal of chromatography. A.

[7]  H. Ji,et al.  Silicon-based microfilters for whole blood cell separation , 2008, Biomedical microdevices.

[8]  T. P. Rivera,et al.  Assisted convective-capillary force assembly of gold colloids in a microfluidic cell: Plasmonic properties of deterministic nanostructures , 2008 .

[9]  S. Bodovitz,et al.  Single cell analysis: the new frontier in 'omics'. , 2010, Trends in biotechnology.

[10]  Brian A. Smith,et al.  Dual-Color-Coded Imaging of Viable Circulating Prostate Carcinoma Cells Reveals Genetic Exchange between Tumor Cells In Vivo, Contributing to Highly Metastatic Phenotypes , 2006, Cell cycle.

[11]  Gang Li,et al.  Highly sensitive enumeration of circulating tumor cells in lung cancer patients using a size-based filtration microfluidic chip. , 2014, Biosensors & bioelectronics.

[12]  T. Becker,et al.  New frontiers in circulating tumor cell analysis: A reference guide for biomolecular profiling toward translational clinical use , 2014, International journal of cancer.

[13]  James N Ingle,et al.  Detection of circulating cytokeratin-positive cells in the blood of breast cancer patients using immunomagnetic enrichment and digital microscopy. , 2002, Clinical cancer research : an official journal of the American Association for Cancer Research.

[14]  Thomas Ragg,et al.  The RIN: an RNA integrity number for assigning integrity values to RNA measurements , 2006, BMC Molecular Biology.

[15]  S. Mocellin,et al.  Molecular detection of circulating tumor cells is an independent prognostic factor in patients with high‐risk cutaneous melanoma , 2004, International journal of cancer.

[16]  A. Dirican,et al.  Prognostic significance of circulating tumor cells and serum CA15-3 levels in metastatic breast cancer, single center experience, preliminary results. , 2013, Asian Pacific journal of cancer prevention : APJCP.

[17]  R. Eils,et al.  Genomic analysis of single cytokeratin-positive cells from bone marrow reveals early mutational events in breast cancer. , 2005, Cancer cell.

[18]  Y. Koh,et al.  Multicolor detection of rare tumor cells in blood using a novel flow cytometry‐based system , 2014, Cytometry. Part A : the journal of the International Society for Analytical Cytology.

[19]  Ruediger Salowsky,et al.  RNA integrity number (RIN) - towards standardization of RNA quality assessment , 2004 .

[20]  Rui Cui,et al.  Detection of circulating tumor cells in breast cancer patients utilizing multiparameter flow cytometry and assessment of the prognosis of patients in different CTCs levels , 2010, Cytometry. Part A : the journal of the International Society for Analytical Cytology.

[21]  James N Turner,et al.  Biochip for separating fetal cells from maternal circulation. , 2007, Journal of chromatography. A.

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

[23]  Sridhar Ramaswamy,et al.  Androgen receptor signaling in circulating tumor cells as a marker of hormonally responsive prostate cancer. , 2012, Cancer discovery.

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

[25]  Mehmet Toner,et al.  Detection of mutations in EGFR in circulating lung-cancer cells. , 2008, The New England journal of medicine.

[26]  Hsueh-Chia Chang,et al.  An integrated dielectrophoretic chip for continuous bioparticle filtering, focusing, sorting, trapping, and detecting. , 2007, Biomicrofluidics.

[27]  Annibale Biggeri,et al.  Correlation of HER2 status between primary tumors and corresponding circulating tumor cells in advanced breast cancer patients , 2009, Breast Cancer Research and Treatment.

[28]  A. Adjei,et al.  Potential Applications for Circulating Tumor Cells Expressing the Insulin-Like Growth Factor-I Receptor , 2007, Clinical Cancer Research.

[29]  D. Moher,et al.  CONSORT 2010 statement: Updated guidelines for reporting parallel group randomised trials , 2010, Journal of pharmacology & pharmacotherapeutics.

[30]  M. Pfaffl,et al.  Comparison of relative mRNA quantification models and the impact of RNA integrity in quantitative real-time RT-PCR , 2006, Biotechnology Letters.

[31]  Y. Kodera,et al.  Chemosensitivity of peritoneal micrometastases as evaluated using a green fluorescence protein (GFP)‐tagged human gastric cancer cell line , 2003, Cancer science.

[32]  Jason P. Gleghorn,et al.  Rare Cell Capture in Microfluidic Devices. , 2011, Chemical engineering science.

[33]  Dino Di Carlo,et al.  Dynamic single-cell analysis for quantitative biology. , 2006, Analytical chemistry.

[34]  Ulrich Keilholz,et al.  Circulating tumor cells: the 'leukemic phase' of solid cancers. , 2006, Trends in molecular medicine.

[35]  C R Cantor,et al.  Biotechnology in the 21st century. , 2000, Trends in biotechnology.

[36]  A. Órfão,et al.  Loss of CD34(+) hematopoietic progenitor cells due to washing can be reduced by the use of fixative-free erythrocyte lysing reagents. , 2000, Journal of immunological methods.

[37]  Joel I. Cohen Harnessing Biotechnology for the Poor: Challenges ahead for capacity, safety and public investment , 2001 .

[38]  I. Cuthill,et al.  Improving Bioscience Research Reporting: The ARRIVE Guidelines for Reporting Animal Research † , 2012, Osteoarthritis and cartilage.