Induction and Assessment of Hypoxia in Glioblastoma Cells In Vitro

Methods Mol Biol. 2018;1741:111-123. doi: 10.1007/978-1-4939-7659-1_9.


To simulate and study the hypoxic microenvironment associated with intracerebral glioma in vivo, simple and reproducible methods are described and discussed for inducing hypoxia or chemical pseudohypoxia in glioma cell cultures and assessing their effects on the expression and nuclear translocation of hypoxia-inducible factor (HIF)-1α, a key transcriptional factor of oxygen homeostasis, by Western blot analysis and immunocytochemistry.

Keywords: Chemical pseudohypoxia; Glioblastoma; Glioma cell lines; Hypoxia cell culture chamber; Hypoxia-inducible factor-1α; Hypoxia-regulated genes; Immunocytochemistry; Prolyl hydroxylases; Tumor hypoxia; Western blot analysis.

Publication types

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

MeSH terms

  • Cell Culture Techniques
  • Cell Hypoxia
  • Cell Line, Tumor
  • Fluorescent Antibody Technique
  • Gene Expression Regulation, Neoplastic
  • Glioblastoma / genetics
  • Glioblastoma / metabolism*
  • Humans
  • Hypoxia / genetics
  • Hypoxia / metabolism*
  • Hypoxia-Inducible Factor 1, alpha Subunit / genetics
  • Hypoxia-Inducible Factor 1, alpha Subunit / metabolism
  • Immunohistochemistry
  • Oxygen Consumption


  • Hypoxia-Inducible Factor 1, alpha Subunit