Identification and Isolation of Cancer Stem Cells Using NANOG-EGFP Reporter System

Methods Mol Biol. 2018:1692:139-148. doi: 10.1007/978-1-4939-7401-6_13.

Abstract

Cancer stem Cells or Cancer Stem-like Cells are thought to be associated with chemoresistance and recurrence in cancer patients following chemotherapy. Developing a method to study these malignant populations is the key to successful development of drug or immunotherapeutic assays. Here, we present a method of identification, isolation of Prostate Cancer Stem Cells (PCSCs) from the DU145 prostate cancer cell line using the NANOG-GFP expression system.

Keywords: Cancer stem cells (CSCs); Fluorescence-activated cell sorting; GFP reporter gene; Lentiviral expression system; NANOG; Prostate cancer (PCa); Sphere formation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Line, Tumor
  • Flow Cytometry
  • Gene Expression Regulation, Neoplastic
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism*
  • Humans
  • Lentivirus / genetics
  • Male
  • Nanog Homeobox Protein / genetics
  • Nanog Homeobox Protein / metabolism*
  • Neoplastic Stem Cells / metabolism*
  • Prostatic Neoplasms / metabolism*

Substances

  • Nanog Homeobox Protein
  • enhanced green fluorescent protein
  • Green Fluorescent Proteins