@article{3076271,
    title = "Algorithmic assessment of cellular senescence in experimental and clinical specimens",
    author = "Kohli, J. and Wang, B. and Brandenburg, S.M. and Basisty, N. and Evangelou, K. and Varela-Eirin, M. and Campisi, J. and Schilling, B. and Gorgoulis, V. and Demaria, M.",
    journal = "Nature Protocols",
    year = "2021",
    volume = "16",
    number = "5",
    pages = "2471-2498",
    publisher = "Lithuanian Nature Research Centre",
    issn = "1754-2189, 1750-2799",
    doi = "10.1038/s41596-021-00505-5",
    keywords = "beta galactosidase;  cell marker;  lipofuscin;  secretory protein;  senescence associated secretory phenotype protein;  unclassified drug;  biological marker, algorithm;  blood sampling;  cell aging;  cell culture;  cell proliferation;  cellular parameters;  clinical protocol;  experimental design;  gene expression;  genetic association;  human;  in vitro study;  in vivo study;  intermethod comparison;  lysosome;  nonhuman;  phenotype;  priority journal;  quantitative analysis;  real time polymerase chain reaction;  Review;  secretory pathway;  senescence associated secretory phenotype;  staining;  tissue preparation;  cytology;  drug effect;  metabolism;  procedures, Algorithms;  Biomarkers;  Cell Proliferation;  Cellular Senescence;  Cytological Techniques;  Humans;  Lysosomes",
    abstract = "The development of genetic tools allowed for the validation of the pro-aging and pro-disease functions of senescent cells in vivo. These discoveries prompted the development of senotherapies—pharmaceutical interventions aimed at interfering with the detrimental effect of senescent cells—that are now entering the clinical stage. However, unequivocal identification and examination of cellular senescence remains highly difficult because of the lack of universal and specific markers. Here, to overcome the limitation of measuring individual markers, we describe a detailed two-phase algorithmic assessment to quantify various senescence-associated parameters in the same specimen. In the first phase, we combine the measurement of lysosomal and proliferative features with the expression of general senescence-associated genes to validate the presence of senescent cells. In the second phase we measure the levels of pro-inflammatory markers for specification of the type of senescence. The protocol can help graduate-level basic scientists to improve the characterization of senescence-associated phenotypes and the identification of specific senescent subtypes. Moreover, it can serve as an important tool for the clinical validation of the role of senescent cells and the effectiveness of anti-senescence therapies. © 2021, The Author(s), under exclusive licence to Springer Nature Limited."
}