Detection of Senescent Cells by Extracellular Markers Using a Flow Cytometry-Based Approach

Methods Mol Biol. 2017:1534:147-153. doi: 10.1007/978-1-4939-6670-7_14.

Abstract

Senescence is a cellular process that is thought to have prognostic and therapeutic relevance in conditions such as cancer, aging, and fibrosis. However, current protocols for identifying senescent cells in vitro and in vivo have several drawbacks. Most markers used lack sufficient specificity and false positives and negatives in common. In addition, classical staining techniques often require lengthy protocols and do not offer objective quantification. Recently, several novel markers of senescence associated with the plasma membrane have been identified. Here, we propose to take advantage of these markers to define a customizable FACS-based protocol to detect senescent cells using antibodies tagged with fluorescence dyes. This method has the advantage of being fast and allowing quantitation. Furthermore, its specificity is increased using several markers simultaneously.

Keywords: Antibodies; Extracellular markers; Flow cytometry; SA-β-Gal; Senescence.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biomarkers*
  • Cells, Cultured
  • Cellular Senescence* / genetics
  • Extracellular Space / metabolism
  • Flow Cytometry / methods*
  • Gene Expression
  • Humans
  • Immunophenotyping
  • beta-Galactosidase / metabolism

Substances

  • Biomarkers
  • beta-Galactosidase