A Multiparametric Assay to Evaluate Senescent Cells

Methods Mol Biol. 2019:1896:107-117. doi: 10.1007/978-1-4939-8931-7_11.

Abstract

Cellular senescence is a permanent growth arrest mechanism triggered by various forms of stress. Senescent cells accumulate in the mammalian organism with age and are present at sites of tissue damage and age related pathologies. However, the characterization of senescence cells in vivo is currently limited and the need for new technologies to detect and monitor the senescence state in vivo has greatly increased. Here we demonstrate the use of the ImageStreamX as a powerful method for detection and quantification of senescent cells at distinct tissues and cell subpopulations. The identification of senescent cells using ImageStreamX enables the use of a combination of several senescence-related markers, together with the commonly used senescence-associated beta-galactosidase assay. These can be combined with the use of other molecular features typical of senescence cells, such as the γH2AX foci, indicating the activation of DNA damage response. This novel method offers a feasible solution to quantify senescent cells in vivo, in a comprehensive manner. Such quantification is necessary in order to understand the role of cellular senescence in aging and disease.

Keywords: Aging; Cellular senescence; DNA damage response; ImageStreamX; SA-β-Gal.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Biological Assay / methods*
  • Biomarkers / analysis*
  • Cellular Senescence*
  • Colon / cytology
  • Colon / metabolism
  • DNA Damage*
  • HMGB1 Protein / metabolism*
  • Histones / metabolism*
  • Lung / cytology
  • Lung / metabolism
  • Mice
  • beta-Galactosidase / metabolism*

Substances

  • Biomarkers
  • HMGB1 Protein
  • HMGB1 protein, mouse
  • Histones
  • gamma-H2AX protein, mouse
  • beta-Galactosidase