Prospective Isolation and Comparison of Human Germinal Matrix and Glioblastoma EGFR + Populations with Stem Cell Properties

Stem Cell Reports. 2017 May 9;8(5):1421-1429. doi: 10.1016/j.stemcr.2017.03.019. Epub 2017 Apr 20.

Abstract

Characterization of non-neoplastic and malignant human stem cell populations in their native state can provide new insights into gliomagenesis. Here we developed a purification strategy to directly isolate EGFR+/- populations from human germinal matrix (GM) and adult subventricular zone autopsy tissues, and from de novo glioblastoma (GBM) resections, enriching for cells capable of binding EGF ligand (LBEGFR+), and uniquely compared their functional and molecular properties. LBEGFR+ populations in both GM and GBM encompassed all sphere-forming cells and displayed proliferative stem cell properties in vitro. In xenografts, LBEGFR+ GBM cells showed robust tumor initiation and progression to high-grade, infiltrative gliomas. Whole-transcriptome sequencing analysis confirmed enrichment of proliferative pathways in both developing and neoplastic freshly isolated EGFR+ populations, and identified both unique and shared sets of genes. The ability to prospectively isolate stem cell populations using native ligand-binding capacity opens new doors onto understanding both normal human development and tumor cell biology.

Keywords: EGFR; FACS; RNA-seq; germinal matrix; glioblastoma; glioma stem cells; neural stem cells; neurosphere; transcriptome; tumor initiation.

MeSH terms

  • Animals
  • Brain Neoplasms / metabolism
  • Brain Neoplasms / pathology*
  • Cell Proliferation*
  • Cell Separation / methods
  • Cells, Cultured
  • ErbB Receptors / genetics
  • ErbB Receptors / metabolism
  • Glioblastoma / metabolism
  • Glioblastoma / pathology*
  • Humans
  • Male
  • Mice, SCID
  • Neoplastic Stem Cells / metabolism
  • Neoplastic Stem Cells / physiology*
  • Neoplastic Stem Cells / transplantation
  • Neural Stem Cells / metabolism
  • Neural Stem Cells / physiology*
  • Primary Cell Culture / methods
  • Transcriptome
  • Xenograft Model Antitumor Assays

Substances

  • EGFR protein, human
  • ErbB Receptors