Detecting the Senescence-Associated Secretory Phenotype (SASP) by High Content Microscopy Analysis

  • Priya Hari
  • Juan Carlos AcostaEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1534)


The diverse arrays of proteins secreted by senescent cells have been described to influence aging and to have both pro-tumorigenic and anti-tumorigenic influences on the surrounding microenvironment. Further characterization of these proteins, known as the senescence-associated secretory phenotype (SASP), and their regulators is required to understand and further manipulate such activities. The use of high-throughput technology allows us to obtain visual and quantitative data on a large number of samples quickly and easily. Not only is this an invaluable tool for conducting large-scale RNAi or compound screenings, but also allows rapid validation of candidates of interest. Here, we describe how we use the Widefield High-Content Analysis Systems to characterize the phenotypes of cells following modulation by potential regulators of Oncogene-Induced Senescence (OIS) by measuring numerous markers of senescence, including the SASP. This approach can be also used to screen for siRNA able to perturb the expression of SASP components during OIS.

Key words

Senescence OIS SASP SMS Senescence secretome High content analysis IL-8 IL-6 IL-1α IL-1β 


  1. 1.
    Pérez-Mancera P, Young A, Narita M (2014) Inside and out: the activities of senescence in cancer. Nat Rev Cancer 14:547–558. doi: 10.1038/nrc3773 CrossRefPubMedGoogle Scholar
  2. 2.
    Salama R, Sadaie M, Hoare M, Narita M (2014) Cellular senescence and its effector programs. Genes Dev 28:99–114. doi: 10.1101/gad.235184.113 CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Muñoz-Espín D, Serrano M (2014) Cellular senescence: from physiology to pathology. Nat Rev Mol Cell Bio 15:482–496. doi: 10.1038/nrm3823 CrossRefGoogle Scholar
  4. 4.
    Kuilman T, Michaloglou C, Vredeveld L et al (2007) Oncogene-induced senescence relayed by an interleukin-dependent inflammatory network. Cell. doi: 10.1016/j.cell.2008.03.039 Google Scholar
  5. 5.
    Acosta J, O’Loghlen A, Banito A et al (2007) Chemokine signaling via the CXCR2 receptor reinforces senescence. Cell. doi: 10.1016/j.cell.2008.03.038 Google Scholar
  6. 6.
    Rodier F, Coppé JP, Patil C et al (2009) Persistent DNA damage signalling triggers senescence-associated inflammatory cytokine secretion. Nat Cell Biol 11:973–979. doi: 10.1038/ncb1909 CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Acosta J, Banito A, Wuestefeld T et al (2013) A complex secretory program orchestrated by the inflammasome controls paracrine senescence. Nat Cell Biol 15:978–990. doi: 10.1038/ncb2784 CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2017

Authors and Affiliations

  1. 1.Edinburgh Cancer Research UK Centre, Institute of Genetics and Molecular MedicineUniversity of EdinburghEdinburghUK

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