Enumeration of Circulating Tumor Cells and Disseminated Tumor Cells in Blood and Bone Marrow by Immunomagnetic Enrichment and Flow Cytometry (IE/FC)

Methods Mol Biol. 2017;1634:203-210. doi: 10.1007/978-1-4939-7144-2_17.

Abstract

Enumerating circulating tumor cells (CTCs) in blood and disseminated tumor cells (DTCs) in bone marrow has shown to be clinically useful, as elevated numbers of these cells predict poor clinical outcomes. Accurate detection and quantification is, however, difficult and technically challenging because CTCs and DTCs are extremely rare. We have developed a novel quantitative detection method for enumeration of CTCs and DTCs. Our approach consists of two steps: (1) EPCAM-based immunomagnetic enrichment followed by (2) flow cytometry (IE/FC). The assay takes approximately 2 h to complete. In addition to tumor cell enumeration, IE/FC offers opportunities for direct isolation of highly pure tumor cells for downstream molecular characterization.

Keywords: Circulating tumor cells; Disseminated tumor cells; Enumeration; Flow cytometry; Immunomagnetic enrichment.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies, Monoclonal / chemistry
  • Antibodies, Monoclonal / metabolism
  • Biomarkers, Tumor / genetics
  • Biomarkers, Tumor / immunology
  • Biomarkers, Tumor / metabolism
  • Bone Marrow Cells / immunology
  • Bone Marrow Cells / metabolism
  • Bone Marrow Cells / pathology
  • Cell Count / instrumentation
  • Cell Count / methods
  • Cell Line, Tumor
  • Epithelial Cell Adhesion Molecule / genetics
  • Epithelial Cell Adhesion Molecule / immunology
  • Epithelial Cell Adhesion Molecule / metabolism
  • Equipment Design
  • Flow Cytometry / instrumentation
  • Flow Cytometry / methods*
  • Fluorescent Dyes / chemistry
  • Humans
  • Immunoconjugates / chemistry
  • Immunomagnetic Separation / instrumentation
  • Immunomagnetic Separation / methods*
  • Leukocyte Common Antigens / genetics
  • Leukocyte Common Antigens / immunology
  • Leukocyte Common Antigens / metabolism
  • Magnetite Nanoparticles / chemistry*
  • Neoplasms / blood
  • Neoplasms / diagnosis*
  • Neoplasms / immunology
  • Neoplasms / pathology
  • Neoplastic Cells, Circulating / immunology
  • Neoplastic Cells, Circulating / metabolism
  • Neoplastic Cells, Circulating / pathology*
  • Protein Binding

Substances

  • Antibodies, Monoclonal
  • Biomarkers, Tumor
  • EPCAM protein, human
  • Epithelial Cell Adhesion Molecule
  • Fluorescent Dyes
  • Immunoconjugates
  • Magnetite Nanoparticles
  • Leukocyte Common Antigens
  • PTPRC protein, human