Supplementary Information files for: Multiparameter flow cytometric detection and quantification of senescent cells in vitro 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-beta-galactosidase 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 b-galactosidase, the expression level of
p16INK4a and cH2AX 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 commonlyused 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