EpCAM-Independent Enrichment and Detection of Viable Circulating Tumor Cells Using the EPISPOT Assay

Methods Mol Biol. 2017:1634:263-276. doi: 10.1007/978-1-4939-7144-2_22.

Abstract

Identification and characterization of circulating tumor cells (CTCs) in peripheral blood can provide information on the direction and the efficacy of treatments. Current techniques such as CellSearch® are limited in differentiating between apoptotic and viable CTCs. In contrast, the fluorescent EPISPOT assay allows for the identification of viable cells by detecting proteins secreted/released/shed by functional single epithelial cancer cells. In addition, as CTCs are rare events, it is required to combine the EPISPOT assay with an enrichment step. In this article, the EPISPOT assay, as well as two technologies for enrichment of viable CTCs, RosetteSep™ and Parsortix™ techniques, will be presented and discussed in detail.

Keywords: Circulating tumor cell enrichment; EPISPOT assay; Functional test; Solid cancers; Viable circulating tumor cells.

MeSH terms

  • Antibodies, Monoclonal / chemistry
  • Antibodies, Monoclonal / metabolism
  • Biomarkers, Tumor / genetics
  • Biomarkers, Tumor / immunology
  • Biomarkers, Tumor / metabolism
  • Cell Count
  • Cell Line, Tumor
  • Cell Separation / instrumentation
  • Cell Separation / methods*
  • Cell Survival
  • Centrifugation, Density Gradient / methods
  • Epithelial Cell Adhesion Molecule
  • Equipment Design*
  • ErbB Receptors / genetics
  • ErbB Receptors / immunology
  • ErbB Receptors / metabolism
  • Ficoll / chemistry
  • Fluorescent Dyes / chemistry
  • Humans
  • Immunoassay
  • Keratin-19 / genetics
  • Keratin-19 / immunology
  • Keratin-19 / metabolism
  • Microfluidic Analytical Techniques / instrumentation*
  • Neoplasms / blood
  • Neoplasms / diagnosis*
  • Neoplasms / immunology
  • Neoplasms / pathology
  • Neoplastic Cells, Circulating / immunology
  • Neoplastic Cells, Circulating / metabolism
  • Neoplastic Cells, Circulating / pathology*
  • Protein Binding
  • Receptor, ErbB-2 / genetics
  • Receptor, ErbB-2 / immunology
  • Receptor, ErbB-2 / metabolism
  • Vascular Endothelial Growth Factor A / genetics
  • Vascular Endothelial Growth Factor A / immunology
  • Vascular Endothelial Growth Factor A / metabolism

Substances

  • Antibodies, Monoclonal
  • Biomarkers, Tumor
  • EPCAM protein, human
  • Epithelial Cell Adhesion Molecule
  • Fluorescent Dyes
  • Keratin-19
  • VEGFA protein, human
  • Vascular Endothelial Growth Factor A
  • Ficoll
  • EGFR protein, human
  • ERBB2 protein, human
  • ErbB Receptors
  • Receptor, ErbB-2