Capture and Genetic Analysis of Circulating Tumor Cells Using a Magnetic Separation Device (Magnetic Sifter)

Methods Mol Biol. 2017;1634:153-162. doi: 10.1007/978-1-4939-7144-2_12.

Abstract

Circulating tumor cells (CTCs) are currently widely studied for their potential application as part of a liquid biopsy. These cells are shed from the primary tumor into the circulation, and are postulated to provide insight into the molecular makeup of the actual tumor in a minimally invasive manner. However, they are extremely rare in blood, with typical concentrations of 1-100 in a milliliter of blood; hence, a need exists for a rapid and high-purity method for isolating CTCs from whole blood. Here, we describe the application of a microfabricated magnetic sifter toward isolation of CTCs from whole blood at volumetric flow rates of 10 mL/h, along with the use of a PDMS-based nanowell system for single-cell gene expression profiling. This method allows rapid isolation of CTCs and subsequent integration with downstream genetic profiling methods for clinical applications such as targeted therapy, therapy monitoring, or further biological studies.

Keywords: Circulating tumor cell; Genetic analysis; Liquid biopsy; Magnetic cell separation; Magnetic sifter; Nanowell array.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Cell Line, Tumor
  • Cell Separation / instrumentation
  • Cell Separation / methods*
  • Cell Size
  • Dimethylpolysiloxanes / chemistry
  • Equipment Design
  • Filtration / instrumentation
  • Filtration / methods*
  • Gene Expression Profiling
  • Gene Expression Regulation, Neoplastic*
  • Humans
  • Lung Neoplasms / blood
  • Lung Neoplasms / diagnosis*
  • Lung Neoplasms / pathology
  • Magnetite Nanoparticles / chemistry
  • Magnets
  • Nanopores
  • Neoplastic Cells, Circulating / metabolism
  • Neoplastic Cells, Circulating / pathology*
  • Nylons / chemistry
  • Rheology
  • Single-Cell Analysis / instrumentation
  • Single-Cell Analysis / methods*

Substances

  • Dimethylpolysiloxanes
  • Magnetite Nanoparticles
  • Nylons
  • poly(dimethylsiloxane)-polyamide copolymer