Simple Detection of Unstained Live Senescent Cells with Imaging Flow Cytometry.

Journal: Cells
Published Date:

Abstract

Cellular senescence is a hallmark of aging and a promising target for therapeutic approaches. The identification of senescent cells requires multiple biomarkers and complex experimental procedures, resulting in increased variability and reduced sensitivity. Here, we propose a simple and broadly applicable imaging flow cytometry (IFC) method. This method is based on measuring autofluorescence and morphological parameters and on applying recent artificial intelligence (AI) and machine learning (ML) tools. We show that the results of this method are superior to those obtained measuring the classical senescence marker, senescence-associated beta-galactosidase (SA-β-Gal). We provide evidence that this method has the potential for diagnostic or prognostic applications as it was able to detect senescence in cardiac pericytes isolated from the hearts of patients affected by end-stage heart failure. We additionally demonstrate that it can be used to quantify senescence "in vivo" and can be used to evaluate the effects of senolytic compounds. We conclude that this method can be used as a simple and fast senescence assay independently of the origin of the cells and the procedure to induce senescence.

Authors

  • Marco Malavolta
    Advanced Technology Center for Aging Research, IRCCS INRCA, 60121 Ancona, Italy.
  • Robertina Giacconi
    Advanced Technology Center for Aging Research, IRCCS INRCA, 60121 Ancona, Italy.
  • Francesco Piacenza
    Advanced Technology Center for Aging Research, IRCCS INRCA, 60121 Ancona, Italy.
  • Sergio Strizzi
    Advanced Technology Center for Aging Research, IRCCS INRCA, 60121 Ancona, Italy.
  • Maurizio Cardelli
    Advanced Technology Center for Aging Research, IRCCS INRCA, 60121 Ancona, Italy.
  • Giorgia Bigossi
    Advanced Technology Center for Aging Research, IRCCS INRCA, 60121 Ancona, Italy.
  • Serena Marcozzi
    Advanced Technology Center for Aging Research, IRCCS INRCA, 60121 Ancona, Italy.
  • Luca Tiano
    Department of Life and Environmental Sciences, Polytechnical University of Marche, 60121 Ancona, Italy.
  • Fabio Marcheggiani
    Department of Life and Environmental Sciences, Polytechnical University of Marche, 60121 Ancona, Italy.
  • Giulia Matacchione
    Department of Clinical and Molecular Sciences, DISCLIMO, Polytechnical University of Marche, 60121 Ancona, Italy.
  • Angelica Giuliani
    Department of Clinical and Molecular Sciences, DISCLIMO, Polytechnical University of Marche, 60121 Ancona, Italy.
  • Fabiola Olivieri
    Department of Clinical and Molecular Sciences, DISCLIMO, Polytechnical University of Marche, 60121 Ancona, Italy.
  • Ilaria Crivellari
    Department of Medicine (DAME), University of Udine, 33100 Udine, Italy.
  • Antonio Paolo Beltrami
    Department of Medicine (DAME), University of Udine, 33100 Udine, Italy.
  • Alessandro Serra
    Luminex B.V., Het Zuiderkruis 1, 5215 MV 's-Hertogenbosch, The Netherlands.
  • Marco Demaria
    European Research Institute for the Biology of Ageing, University Medical Center Groningen, University of Groningen, The Netherlands.
  • Mauro Provinciali
    Advanced Technology Center for Aging Research, IRCCS INRCA, 60121 Ancona, Italy.