Adewoye2020_Article_MultiparameterFlowCytometricDe.pdf (1.2 MB)

Multiparameter flow cytometric detection and quantification of senescent cells in vitro

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posted on 04.08.2020, 10:02 by Adeolu Badi Adewoye, Dimitris Tampakis, Antonia Follenzi, Alexandra Stolzing
It has been over half a century since cellular senescence was first noted and characterized, and yet no consensus senescent marker has been reliably established. This challenge is compounded by the complexity and heterogenic phenotypes of senescent cells. This necessitates the use of multiple biomarkers to confidently characterise senescent cells. Despite cytochemical staining of senescence associated-betagalactosidase being a single marker approach, as well as being time and labour-intensive, it remains the most popular detection method. We have developed an alternative flow cytometry-based method that simultaneously quantifies multiple senescence markers at a single-cell resolution. In this study, we applied this assay to the quantification of both replicative and induced senescent primary cells. Using this assay, we were able to quantify the activity level of SA β-galactosidase, the expression level of p16INK4a and γH2AX in these cell populations. Our results show this flow cytometric approach to be sensitive, robust, and consistent in discriminating senescent cells in different cell senescence models. A strong positive correlation between these commonly- used senescence markers was demonstrated. The method described in this paper can easily be scaled up to accommodate high-throughput screening of senescent cells in applications such as therapeutic cell preparation, and in therapy-induced senescence following cancer treatment.

Funding

European Union’s Horizon 2020 research and innovation programme under Grant Agreement No. 667421.

History

School

  • Mechanical, Electrical and Manufacturing Engineering

Published in

Biogerontology

Volume

21

Issue

6

Pages

773 - 786

Publisher

Springer

Version

VoR (Version of Record)

Rights holder

© The Authors

Publisher statement

This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

Acceptance date

30/07/2020

Publication date

2020-08-10

Copyright date

2020

ISSN

1389-5729

eISSN

1573-6768

Language

en

Depositor

Prof Alexandra Stolzing. Deposit date: 3 August 2020

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