Robust multiparametric assessment of cellular senescence

Clara Correia-Melo, Diana Jurk, João F. Passos

Research output: Chapter in Book/Report/Conference proceedingChapter

6 Scopus citations

Abstract

Cellular senescence, the irreversible loss of replicative capacity, is both a tumor suppressor mechanism and a contributor to the age-related loss of tissue function. However, the role of cellular senescence in vivo has been unclear, mostly because of the absence of cellular markers specific enough to identify the state (senescent or proliferating) of individual cells in tissues. Recently, we have tested the robustness of multiple senescence candidate markers by comparing them to a dynamic stimulation model, which estimates the fraction of senescent cells with high precision. We found that the absence of the proliferation markers Ki67 and PCNA combined with high density DNA damage foci (>5 γH2AX foci per nucleus) was the best quantitative indicator of cellular senescence. In this chapter, we describe protocols for the dual immuno fluorescence-based quantification of Ki67/PCNA and γH2AX in both fixed cells and paraf finembedded tissues.

Original languageEnglish (US)
Title of host publicationCell Senescence
Subtitle of host publicationMethods and Protocols
PublisherHumana Press Inc.
Pages409-419
Number of pages11
ISBN (Print)9781627032384
DOIs
StatePublished - 2013

Publication series

NameMethods in Molecular Biology
Volume965
ISSN (Print)1064-3745

Keywords

  • Cellular senescence
  • Immunofluorescence
  • Ki-67
  • PCNA
  • γH2AX

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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