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

Mark Jesus M Magbanua; Tulasi I Solanki; Andrea D Ordonez; Feng Hsiao; John W Park

doi:10.1007/978-1-4939-7144-2_17

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.

Authors

Mark Jesus M Magbanua¹, Tulasi I Solanki², Andrea D Ordonez², Feng Hsiao², John W Park²

Affiliations

¹ Division of Hematology Oncology, Helen Diller Family Comprehensive Cancer Center, University of California-San Francisco, Box 1387, 2340 Sutter St., S471, San Francisco, CA, 94115, USA. mark.magbanua@ucsf.edu.
² Division of Hematology Oncology, Helen Diller Family Comprehensive Cancer Center, University of California-San Francisco, Box 1387, 2340 Sutter St., S471, San Francisco, CA, 94115, USA.

PMID: 28819853
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