STAR Protocols (Mar 2024)
One-step rapid tracking and isolation of senescent cells in cellular systems, tissues, or animal models via GLF16
- Sophia Magkouta,
- Dimitris Veroutis,
- Athanasios Pousias,
- Angelos Papaspyropoulos,
- Kety Giannetti,
- Natassa Pippa,
- Nikolaos Lougiakis,
- Konstantinos Kambas,
- Nefeli Lagopati,
- Aikaterini Polyzou,
- Maria Georgiou,
- Maria Chountoulesi,
- Stergios Pispas,
- Spyros Foutadakis,
- Efthymios Kyrodimos,
- Nicole Pouli,
- Panagiotis Marakos,
- Athanassios Kotsinas,
- Panayotis Verginis,
- Dimitrios Valakos,
- Giannis Vatsellas,
- Russell Petty,
- Dimitris Thanos,
- Marco Demaria,
- Konstantinos Evangelou,
- Raffaella Di Micco,
- Vassilis G. Gorgoulis
Affiliations
- Sophia Magkouta
- Molecular Carcinogenesis Group, Department of Histology and Embryology, Medical School, National and Kapodistrian University of Athens, 11527 Athens, Greece; Marianthi Simou and G.P. Livanos Labs, 1st Department of Critical Care and Pulmonary Services, School of Medicine, National & Kapodistrian University of Athens, ‘‘Evangelismos’’ Hospital, 10676 Athens, Greece
- Dimitris Veroutis
- Molecular Carcinogenesis Group, Department of Histology and Embryology, Medical School, National and Kapodistrian University of Athens, 11527 Athens, Greece; Biomedical Research Foundation, Academy of Athens, 11527 Athens, Greece
- Athanasios Pousias
- Department of Pharmacy, Division of Pharmaceutical Chemistry, National and Kapodistrian University of Athens, Panepistimiopolis, Zografou, 15771 Athens, Greece
- Angelos Papaspyropoulos
- Molecular Carcinogenesis Group, Department of Histology and Embryology, Medical School, National and Kapodistrian University of Athens, 11527 Athens, Greece; Biomedical Research Foundation, Academy of Athens, 11527 Athens, Greece
- Kety Giannetti
- San Raffaele Telethon Institute for Gene Therapy (SR-TIGET), IRCCS San Raffaele Scientific Institute, 20132 Milan, Italy
- Natassa Pippa
- Section of Pharmaceutical Technology, Department of Pharmacy, School of Health Sciences, National and Kapodistrian University of Athens, Panepistimioupolis Zografou, 15771 Athens, Greece; Theoretical and Physical Chemistry Institute, National Hellenic Research Foundation, 11635 Athens, Greece
- Nikolaos Lougiakis
- Department of Pharmacy, Division of Pharmaceutical Chemistry, National and Kapodistrian University of Athens, Panepistimiopolis, Zografou, 15771 Athens, Greece
- Konstantinos Kambas
- Hellenic Pasteur Institute, 11521 Athens, Greece
- Nefeli Lagopati
- Molecular Carcinogenesis Group, Department of Histology and Embryology, Medical School, National and Kapodistrian University of Athens, 11527 Athens, Greece; Biomedical Research Foundation, Academy of Athens, 11527 Athens, Greece; Laboratory of Biology, Department of Basic Medical Sciences, Medical School, National and Kapodistrian University of Athens, 11527 Athens, Greece
- Aikaterini Polyzou
- Molecular Carcinogenesis Group, Department of Histology and Embryology, Medical School, National and Kapodistrian University of Athens, 11527 Athens, Greece
- Maria Georgiou
- Department of Pharmacy, Division of Pharmaceutical Chemistry, National and Kapodistrian University of Athens, Panepistimiopolis, Zografou, 15771 Athens, Greece
- Maria Chountoulesi
- Section of Pharmaceutical Technology, Department of Pharmacy, School of Health Sciences, National and Kapodistrian University of Athens, Panepistimioupolis Zografou, 15771 Athens, Greece
- Stergios Pispas
- Theoretical and Physical Chemistry Institute, National Hellenic Research Foundation, 11635 Athens, Greece
- Spyros Foutadakis
- Biomedical Research Foundation, Academy of Athens, 11527 Athens, Greece
- Efthymios Kyrodimos
- First ENT Department, Hippocration Hospital, National Kapodistrian University of Athens, 11527 Athens, GR, Greece
- Nicole Pouli
- Department of Pharmacy, Division of Pharmaceutical Chemistry, National and Kapodistrian University of Athens, Panepistimiopolis, Zografou, 15771 Athens, Greece
- Panagiotis Marakos
- Department of Pharmacy, Division of Pharmaceutical Chemistry, National and Kapodistrian University of Athens, Panepistimiopolis, Zografou, 15771 Athens, Greece
- Athanassios Kotsinas
- Molecular Carcinogenesis Group, Department of Histology and Embryology, Medical School, National and Kapodistrian University of Athens, 11527 Athens, Greece
- Panayotis Verginis
- Biomedical Research Foundation, Academy of Athens, 11527 Athens, Greece; Laboratory of Immune Regulation and Tolerance, Division of Basic Sciences, University of Crete Medical School, 70013 Heraklion, Greece; Institute of Molecular Biology and Biotechnology, Foundation for Research and Technology, 70013 Heraklion, Greece
- Dimitrios Valakos
- Biomedical Research Foundation, Academy of Athens, 11527 Athens, Greece
- Giannis Vatsellas
- Greek Genome Center, Biomedical Research Foundation, Academy of Athens, 11527 Athens, Greece
- Russell Petty
- Ninewells Hospital and Medical School, University of Dundee, Dundee DD19SY, UK
- Dimitris Thanos
- Biomedical Research Foundation, Academy of Athens, 11527 Athens, Greece; Greek Genome Center, Biomedical Research Foundation, Academy of Athens, 11527 Athens, Greece
- Marco Demaria
- European Research Institute for the Biology of Ageing (ERIBA), University Medical Center Groningen, Groningen 9713 AV, The Netherlands
- Konstantinos Evangelou
- Molecular Carcinogenesis Group, Department of Histology and Embryology, Medical School, National and Kapodistrian University of Athens, 11527 Athens, Greece
- Raffaella Di Micco
- San Raffaele Telethon Institute for Gene Therapy (SR-TIGET), IRCCS San Raffaele Scientific Institute, 20132 Milan, Italy
- Vassilis G. Gorgoulis
- Molecular Carcinogenesis Group, Department of Histology and Embryology, Medical School, National and Kapodistrian University of Athens, 11527 Athens, Greece; Biomedical Research Foundation, Academy of Athens, 11527 Athens, Greece; Ninewells Hospital and Medical School, University of Dundee, Dundee DD19SY, UK; Faculty Institute for Cancer Sciences, Manchester Academic Health Sciences Centre, University of Manchester, Manchester M20 4GJ, UK; Faculty of Health and Medical Sciences, University of Surrey, Surrey GU2 7YH, UK; Corresponding author
- Journal volume & issue
-
Vol. 5,
no. 1
p. 102929
Abstract
Summary: Identification and isolation of senescent cells is challenging, rendering their detailed analysis an unmet need. We describe a precise one-step protocol to fluorescently label senescent cells, for flow cytometry and fluorescence microscopy, implementing a fluorophore-conjugated Sudan Black-B analog, GLF16. Also, a micelle-based approach allows identification of senescent cells in vivo and in vitro, enabling live-cell sorting for downstream analyses and live in vivo tracking. Our protocols are applicable to cellular systems, tissues, or animal models where senescence is present.For complete details on the use and execution of this protocol, please refer to Magkouta et al.1
